Journal: Journal of Cellular and Molecular Medicine

Article Title: Simvastatin preparations promote PDGF‐BB secretion to repair LPS‐induced endothelial injury through the PDGFRβ/PI3K/Akt/IQGAP1 signalling pathway

doi: 10.1111/jcmm.14709

Figure Lengend Snippet: The effects of SV preparations on PDGFRβ phosphorylation, Akt phosphorylation, IQGAP1 expression and PDGF‐BB levels in supernatants of HUVECs cultures treated with LPS. (A‐C) The ratios of p‐PDGFRβ to total PDGFRβ protein and p‐Akt to total Akt protein were remarkably decreased in the LPS group compared with control group and SV/SV‐NP groups (all P < .01), the ratio of p‐PDGFRβ to total PDGFRβ in SV‐NP group was higher than that in SV group ( P < .01), and there is no significant difference in Akt phosphorylation between SV and SV‐NP groups. (A, D) The relative protein levels are expressed as the ratio of the target protein to glyceraldehyde 3‐phosphate dehydrogenase (GAPDH), the levels of IQGAP1 were remarkably down‐regulated in the LPS group compared with control group and SV/SV‐NP groups (all P < .01), and the IQGAP1 protein level in SV‐NP group was higher than that in SV group ( P < .05). (E) The levels of PDGF‐BB in supernatants of the cultures were increased in the SV and SV‐NP groups compared with LPS group (all P < .01), and the PDGF‐BB level was higher in SV‐NP group than SV group ( P < .01). * P < .05 vs control group, ** P < .01 vs control group, # P < .05, ## P < .01

Article Snippet: For immunoprecipitation, 20 μL of protein A magnetic beads (#161‐4013, BIO‐RAD, USA) was precipitated with IQGAP1 antibody (1:100, #29016, Cell Signal Technology, USA) for 1 hour at room temperature.

Techniques: Phospho-proteomics, Expressing, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Simvastatin preparations promote PDGF‐BB secretion to repair LPS‐induced endothelial injury through the PDGFRβ/PI3K/Akt/IQGAP1 signalling pathway

doi: 10.1111/jcmm.14709

Figure Lengend Snippet: The effects of IQGAP1 knockdown on TEER, monolayer permeability and cell migration of LPS‐injured HUVECs treated with SV preparations. (A) The TEER of different simvastatin preparations treatments on LPS‐injured HUVECs after IQGAP1 knockdown at multiple time‐points. (B) The TEER at 24 hours was significantly decreased in LPS + IQGAP1 siRNA group compared with LPS group ( P < .05) and decreased in SV/SV‐NP + IQGAP1 siRNA groups compared with SV/SV‐NP groups (all P < .01), and the TEER at 24 hours in SV/SV‐NP + IQGAP1 siRNA groups was higher than those in LPS + IQGAP1 siRNA group (all P < .01). (C) EB concentrations in the lower chambers were significantly increased in LPS + IQGAP1 siRNA group compared with LPS group ( P < .01) and increased in SV/SV‐NP + IQGAP1 siRNA groups compared with SV/SV‐NP groups (all P < .05), and the EB concentrations in SV/SV‐NP + IQGAP1 siRNA groups were lower than those in LPS + IQGAP1 siRNA group (all P < .01). (D, E) Cell confluence rates at 24 hours were significantly decreased in the LPS + IQGAP1 siRNA group compared with the LPS group ( P < .01) and decreased in SV/SV‐NP + IQGAP1 siRNA groups compared with SV/SV‐NP groups (all P < .01), the confluence rates in SV/SV‐NP + IQGAP1 siRNA groups were higher than those in LPS + IQGAP1 siRNA group (all P < .01). (F, G) IQGAP1 protein level was down‐regulated in the LPS + IQGAP1 siRNA group compared with the LPS group ( P < .05) and decreased in SV/SV‐NP + IQGAP1 siRNA groups compared with SV/SV‐NP groups (all P < .01), and IQGAP1 protein level in SV/SV‐NP + IQGAP1 siRNA groups was higher than those in LPS + IQGAP1 siRNA group (all P < .01). The negative control siRNA has no significant effect on TEER, monolayer permeability, cell migration (Figure ), PDGFRβ phosphorylation, Akt phosphorylation, IQGAP1 expression and PDGF‐BB in supernatants of the cultures of HUVECs (Figure ). * P < .05 vs LPS group, ** P < .01 vs LPS group, # P < .05, ## P < .01

Article Snippet: For immunoprecipitation, 20 μL of protein A magnetic beads (#161‐4013, BIO‐RAD, USA) was precipitated with IQGAP1 antibody (1:100, #29016, Cell Signal Technology, USA) for 1 hour at room temperature.

Techniques: Knockdown, Permeability, Migration, Negative Control, Phospho-proteomics, Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Simvastatin preparations promote PDGF‐BB secretion to repair LPS‐induced endothelial injury through the PDGFRβ/PI3K/Akt/IQGAP1 signalling pathway

doi: 10.1111/jcmm.14709

Figure Lengend Snippet: The effects of PI3K/Akt inhibition on PDGFRβ phosphorylation, IQGAP1 expression and PDGF‐BB in supernatants of cultures of LPS‐injured HUVECs treated with SV preparations. (A, B) The ratio of p‐PDGFRβ to total PDGFRβ protein was remarkably decreased in the SV/SV‐NP + LY294002 groups compared with the SV/SV‐NP groups (SV + LY294002: P < .05, SV‐NP + LY294002: P < .01). (A, C) The ratio of p‐Akt to total Akt protein was remarkably decreased in the SV/SV‐NP + LY294002 groups compared with the SV/SV‐NP groups (all P < .01). (A, D, E) The IQGAP1 protein level and PDGF‐BB in supernatants of the cultures in the SV/SV‐NP + LY294002 groups were remarkably down‐regulated than those in the SV/SV‐NP groups (all P < .01). The LY294002 has no significant effect on PDGFRβ phosphorylation, Akt phosphorylation, IQGAP1 expression and PDGF‐BB in supernatants of the cultures of HUVECs (Figure ). * P < .05 vs SV + LPS group, ** P < .01 vs SV + LPS group, ## P < .01

Article Snippet: For immunoprecipitation, 20 μL of protein A magnetic beads (#161‐4013, BIO‐RAD, USA) was precipitated with IQGAP1 antibody (1:100, #29016, Cell Signal Technology, USA) for 1 hour at room temperature.

Techniques: Inhibition, Phospho-proteomics, Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Simvastatin preparations promote PDGF‐BB secretion to repair LPS‐induced endothelial injury through the PDGFRβ/PI3K/Akt/IQGAP1 signalling pathway

doi: 10.1111/jcmm.14709

Figure Lengend Snippet: The effects of PDGFRβ inhibition on TEER, monolayer permeability, cell migration and the PDGFRβ/PI3K/Akt/IQGAP1 pathway activity of LPS‐injured HUVECs treated with SV preparations. (A) The TEER of different simvastatin preparations treatments on LPS‐injured HUVECs after PDGFR inhibition at multiple time‐points. (B) The TEER at 24 hours was significantly decreased in the SV/SV‐NP + Imatinib groups compared with the SV/SV‐NP groups (all P < .01). (C) EB concentrations in the lower chambers were increased in SV/SV‐NP + Imatinib groups compared with SV/SV‐NP groups (all P < .01). (D, E) Cell confluence rates at 24 hours were significantly decreased in the SV/SV‐NP + Imatinib groups compared to the SV/SV‐NP groups (all P < .01). (F‐H) The ratios of p‐PDGFRβ to total PDGFRβ protein and p‐Akt to total Akt protein were remarkably decreased in the SV/SV‐NP + Imatinib groups compared with the SV/SV‐NP groups (all P < .01). (F, I, J) IQGAP1 protein level and PDGF‐BB in supernatants of the cultures were remarkably decreased in the SV/SV‐NP + Imatinib groups compared with the SV/SV‐NP groups (all P < .01). The Imatinib has no significant effect on TEER, monolayer permeability, cell migration and PDGFRβ/PI3K/Akt/IQGAP1 pathway activity of HUVECs (Figure , 2). * P < .05 vs SV + LPS group, ** P < .01 vs SV + LPS group, ## P < .01

Article Snippet: For immunoprecipitation, 20 μL of protein A magnetic beads (#161‐4013, BIO‐RAD, USA) was precipitated with IQGAP1 antibody (1:100, #29016, Cell Signal Technology, USA) for 1 hour at room temperature.

Techniques: Inhibition, Permeability, Migration, Activity Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Simvastatin preparations promote PDGF‐BB secretion to repair LPS‐induced endothelial injury through the PDGFRβ/PI3K/Akt/IQGAP1 signalling pathway

doi: 10.1111/jcmm.14709

Figure Lengend Snippet: The effects of SV preparations on the PDGFRβ/IQGAP1 binding ability of HUVECs treated with LPS and the effects of PDGFRβ/PI3K/Akt inhibition on the PDGFRβ/IQGAP1‐binding ability of LPS‐injured HUVECs treated with SV preparations. (A, C) Detection of PDGFRβ/IQGAP1 physical association by immunoprecipitation. The PDGFRβ/IQGAP1 binding was remarkably decreased in LPS group compared with control group ( P < .05) and SV/SV‐NP groups (SV: P < .05, SV‐NP: P < .01). (B, D) PDGFRβ/IQGAP1 binding ability was significantly decreased in LY294002 + SV/SV‐NP groups and Imatinib + SV/SV‐NP groups compared with the SV/SV‐NP groups (all P < .01). The negative control siRNA, LY294002 and imatinib have no significant effect on PDGFRβ/IQGAP1 binding. Normal IgG was used as a negative control (Figure ). * P < .05 vs control/SV group, ** P < .01 vs control/SV group, # P < .05, ## P < .01

Article Snippet: For immunoprecipitation, 20 μL of protein A magnetic beads (#161‐4013, BIO‐RAD, USA) was precipitated with IQGAP1 antibody (1:100, #29016, Cell Signal Technology, USA) for 1 hour at room temperature.

Techniques: Binding Assay, Inhibition, Immunoprecipitation, Control, Negative Control