Figures & data
Figure 1. Structural information and safety assessment of NGR1. (A) Structural information of NGR1. (B-F) Different parameters of animal after receiving an intraperitoneal injection of NGR1 (30 mg/kg, once per day for 3 d) after 7 d. B: Body weight change rate (7 d). C: Liver weight (% of total body weight). D: Serum ALT levels. E: Serum AST levels. F: Food intake (g/5 mice after 7 d). (G,H) The cell viability of IMVECs treated with different concentrations of NGR1 for 30 min through different assays.G: MTT asay, H:CCK-8 assay. NS, p > 0.05 compared with the control group.
Figure 2. Effects of NGR1 on survival and intestinal function of septic mice. (A) Survival rate and survival time (n = 15). (B) HE staining of intestinal villi to observe gross morphology (400×). (C) Speckle tomography images of mesenteric perfusion and related statistics. (D) FITC-BSA leakage of mesenteric microveins (n = 8). Data are presented as mean ± standard deviation. a, p < 0.05 compared with the control group; b, p < 0.05 compared with the CLP group.
Figure 3. Effects of NGR1 on mitochondrial quality in LPS-induced IMVECs. (A) Representative confocal images of IMVEC mitochondrial morphology in each group (bar = 25 μm) and analysis of mitochondrial skeletons using Image J software. (B) Representative confocal images of mitochondrial membrane potential (ΔΨm) of IMVECs, which were labeled with JC-1 monomer (green fluorescent probe) and JC-1 aggregate (red fluorescent probe) (bar = 25 μm). (C) Representative confocal images of ROS fluorescence intensity in IMVECs (bar = 50 μm). (D) Statistical analysis of ΔΨm and ROS in IMVECs (n = 5). Data are presented as mean ± standard deviation. a, p < 0.05 compared with the control group; b, p < 0.05 compared with the LPS group.
Figure 4. Targeting effect of NGR1 on Drp1, verified using small-molecule microarray chips. (A) Flow chart of small-molecule microarray chips used for screening interactions with Drp1. (B) Display of chip results. From left to right are the chip scan of experimental sample Drp1-biotin and global chip scan of control sample 6× His-tag biotin; chip results are analysed in the table below. (C) From left to right, the structural formula of NGR1, overall structural diagram of NGR1 docking to Drp1 (yellow portion is Drp1, pink portion is small molecule NGR1), and local diagram of the specific interaction sites.
Figure 5. Effects of NGR1 on Drp1 expression in subcellular fractions of IMVECs. Western blotting results showing the effect of NGR1 treatment on the expression of (A) total Drp1, (B) mitochondrial Drp1, and (C) cytoplasmic Drp1 in IMVECs under sepsis (n = 5). Data are presented as mean ± standard deviation. a, p < 0.05 compared with the control group, b, p < 0.05 compared with the LPS group. NS, p > 0.05 compared with the control group.
Data availability statement
The datasets of this study can be found at https://www.jianguoyun.com/p/DVeURM0QtMvLChjY0fUEIAA.