Figure 1.
Experimental protocol.
Figure 2.
Liver IPC improved hepatic graft function following LT in rats.
(a) Representative hepatic graft histopathology stained with hematoxylin and eosin (H&E, 200×). (b) Changes of liver enzymes (ALT and AST) in the serum of rats following LT.
Figure 3.
Liver IPC improved intestinal barrier function and inflammation following LT in rats.
(a) Representative intestinal mucosal ultrastructure obtained by transmission electron microscopy (TEM) in rats following LT. MV: microvilli, TJ: tight junction. (b) Serum endotoxin was detected in the different groups. (c) The content of sIgA in wet feces was calculated by ELISA and expressed as nanogram (ng) per gram wet feces in the different groups. (d) Serum TNF-α was determined by ELISA in the different groups. *p<0.05, **p<0.01, ***p<0.001.
Figure 4.
Quantitative analysis of characteristic bacteria in feces.
Log10 copies/g: log10 no. of 16S rDNA gene copies per gram feces (wet weight). On the level of bacterial genus, specific bacterial populations were determined by real-time qPCR in the different groups: (a) Bifidobacterium spp., (b) Clostridium cluster XIVab and (c) Clostridium clusters XI. Comparisons of specific bacteria among the NC group, DL group and Liver-IPC group were emphasized. Statistical differences were calculated by one-way ANOVA. *p<0.05, **p<0.01, ***p<0.001.
Figure 5.
Liver IPC improved intestinal microbiota following LT in rats shown in DGGE profiles.
(a) DGGE profiles of fecal bacteria in rats from the different groups. Sample numbers above lanes indicated the different rats from the different groups. Marker lane was used for gel to gel comparison. Each band represents a bacterial clone. Band numbers (corresponding to Figure 7 band classes) indicated the position of bands excised for sequence analyses (e.g. ‘“20”’ means band 20). (b) Intestinal microbial diversity comparison (Shannon’s diversity index). (c) Species richness comparison. (d) Shannon’s evenness index comparison. *p<0.05, **p<0.01.
Figure 6.
Cluster analysis of DGGE profiles obtained with universal primers V3 using Dice’s coefficient and UPGMA.
(a) Cluster analysis of DGGE profiles from the different groups. Metric scale denotes the degree of similarity. (b) Multidimensional scaling (MDS) analysis of the cluster shown in (a). The plot is an optimized three-dimensional representation of the similarity matrix obtained from BioNumerics software, and the x-, y-, and z-axes separately represent three different dimension: Dim 1, Dim 2, and Dim 3. Euclidean distance between two points reflects similarity. (c) Principal components analysis (PCA) of fecal microbiota based on DGGE fingerprinting shown in (a). It reorients the plot to maximize the variation among lanes along the first three principal components (the contributions 21.9, 20.1 and 13.3, respectively) obtained from BioNumerics software.
Figure 7.
Phylogenetic tree of sequences using the neighbor-joining method (a) and key bands of structural shift of intestinal microbiota (b and c) from DGGE profiles.
(a) Phylogenetic tree of sequences from DGGE profiles. The fragment sequences were named for their positions in gels using the band-matching tool with BioNumerics software version 6.01 (Applied Maths). The numbers shown in Figure 5 are in coincidence with the numbers shown in the brackets in the figure. 23 band classes without labels showed little variation in intensity among the different groups. Compared to the NC group, 8 band classes labeled with the black square showed a decrease in intensity, while 8 band classes labeled with the black triangle showed an increase in the DL group; Compared to the DL group, 3 band classes labeled with the black diamond showed a decrease in intensity, while 8 band classes labeled with the black spot showed an increase in the Liver IPC group. The plot was obtained from MEGA5 software (http://en.wikipedia.org/wiki/MEGA,_Molecular_Evolutionary_Genetics_Analysis). (b) The intensities of 3 key bands (band class 48.8, 53.3 and 70.6) were increased in the DL group and then decreased after liver IPC, and band class 45.0 also showed this trend. (c) The intensities of 4 key bands (band class 85.2, 88.0, 13.9 and 42.7) were decreased in the DL group and then increased after liver IPC. Both band classes 73.5 and 78.0 also showed this trend.