Research Paper Volume 12, Issue 2 pp 1397—1416

Activated Drp1-mediated mitochondrial ROS influence the gut microbiome and intestinal barrier after hemorrhagic shock

Figure 2. The effects of Drp1 on intestinal barrier function after shock. (A) The genotype identification results of Drp1 KO and WT mice. The genotype of Drp1 KO mice (blue dot, n=8) is Drp1 +/- (double bands at 1115 bp and 472 bp), and the genotype of WT mice (red dot, n=8) is Drp1 +/+ (single band at 472 bp). (B) Drp1 mRNA expression in colon tissues of HS_WT group and HS_Drp1 KO group detected by QRT-PCR (n=8 mice/ group). (C) Drp1 protein expression in colon tissues of HS_WT group and HS_Drp1 KO group detected by Western Blotting (n=8 mice/ group). (D) The mitochondrial morphology and intestinal epithelial tight junctions observed by electronic microscopy (TEM). MT, mitochondria (black arrows refer to vacuolated cristae structure); TJ, tight junctions (white arrows). The boundaries of mitochondria are depicted and the lengths as well as cristae density of all mitochondria in ROI (region of interest) are calculated by Image J software. The mitochondrial number in ROI: 2 in N_WT group, 10 in HS_WT group and 6 in HS_Drp1 KO group. (Bar, 500nm). (E) The intestinal villus morphology observed by immunohistochemistry (400X). (F) The plasma D-lactate and endotoxin concentration in each group (n=8 mice/ group). N_WT, WT mice in normal condition; HS_WT, WT mice in hemorrhagic shock condition; HS_Drp1 KO, Drp1 KO mice in hemorrhagic shock condition. a represents p < 0.05 compared with N_WT group; b represents p < 0.05 compared with HS_WT group.