Fig. 2.

Pericyte underwent ferroptosis and inhibition of ferroptosis alleviated sepsis-related pulmonary vascular leakage. (A) U-cell score of different programmed cell death (PCD) of pericytes in mice following sepsis. (B) Volcano plot of differentially expressed genes between control groups and LPS treated groups. (C) KEGG pathway (upregulated) of differentially expressed genes between control groups and LPS treated groups. (D) Cell viability of LPS-stimulated pericytes pretreated with inhibitors of different PCD. (E) The levels of lipid peroxidation, ferrous iron, GSH and GSH/GSSG ratio of LPS-stimulated pericytes pretreated with Fer-1. (F) FITC-BSA leakage in endothelial cells. (G) Flow cytometry analyses of ferroptosis of CD31⁻CD140b⁺ cells (pericytes) in the lung of mice. (n = 8/group) (H) Western blotting analysis of pericyte marker (PDGFR-β and NG2) in the lung tissues. (n = 3/group) (I) Laser confocal microscopy analysis of the pulmonary networks. (NG2, green, pericyte marker; CD31, red, endothelial cell marker. Bar = 50 μm). (n = 8/group) (J) Representative images of Evens-Blue leakage in the lung tissues. (n = 8/group) (K) Representative TEM images of tight junctions in pulmonary vessels. (Green arrow indicate the tight junction. Bar = 1 μm) (L) Wet weight to Dry weight ratio of lung in septic rats treated by Fer-1. (n = 8/group) (M) Inflammatory factors levels (IL-6 and TNF-α) in serum. (n = 8/group) (N) Survival curves of rats undergoing sham surgery, CLP and Fer-1 pretreatment. (n = 16/group)

Data were shown as mean ± SD. ∗∗p < 0.01, ∗∗∗p < 0.001. Statistical significance was determined by one-way ANOVA or two-sided Student's-test as appropriate. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)