Fig. 5. The effects of UC-MSCs on upregulating autophagy in the livers of mice with I/R injury depends on the activation of AMPKα.

To detect the activation of AMPK and its downstream autophagy-related signaling pathway, mice with liver I/R injury and treated with PBS or UC-MSCs were killed at 6 and 24 h after reperfusion. In addition, to investigate the role of AMPK activation, model mice treated with UC-MSCs and dorsomorphin were killed 6 h after reperfusion. a Western blot analysis revealed the expression of AMPK, p-AMPK, mTOR, p-mTOR, ULK and p-ULK in liver tissues from each group. The average intensities of the band in the western blots were quantified using GAPDH as an internal reference. b TEM was used to determine the number of autophagic vacuoles. The data are presented as the means ± SEMs (n = 5/group). c The hepatic histology of each experimental group was obtained by H&E staining (magnification ×100), and the degree of liver injury was determined using Suzike’s injury score. The data are presented as the means ± SEMs (n = 5/group). d Representative liver sections from each group stained with caspase-3 were obtained (magnification ×200). The IHC staining of caspase-3 in liver tissues was semiquantified (n = 5). The data are presented as the means ± SEMs (n = 5/group). e Representative liver sections from each experimental group stained with DHE and DAPI were obtained to determine the level of total ROS. The data are presented as the means ± SEMs (n = 5/group). f Representative liver sections from each experimental group stained with MitoSOX Red and DAPI were obtained to determine the level of mtROS. The data are presented as the means ± SEMs (n = 5/group). *p < 0.05, **p < 0.01, ***p < 0.001 (all p-values were obtained by one-way ANOVA).