Journal: International Journal of Medical Sciences
Article Title: Stem cells ameliorate neurotrauma-induced visual disturbances and retinal degeneration via broad normalization of β-catenin-related signaling
doi: 10.7150/ijms.123975
Figure Lengend Snippet: MSC therapy enhances growth factor expression and receptor signaling in SI-injured R28 cells and TBI rats. (A) Representative growth factor antibody array images from conditioned media of R28 cells subjected to stretch injury (SI), with or without MSC coculture. Increased secretion of HGF, IGFBP-4, IGFBP-6, and VEGF was observed in the SI+MSC group. Positive controls are marked in red boxes; significantly regulated proteins are highlighted in blue. Quantitative analysis of mean pixel density is shown. (B) ELISA quantification of serum VEGF, HGF, IGFBP-4, and IGFBP-6 levels in TBI rats. TBI reduced systemic levels of these growth factors, while MSC treatment significantly restored them (except IGFBP-6). (C) ELISA quantification of VEGF, HGF, IGFBP-4, and IGFBP-6 in whole eyeball lysates (OD and OS). Due to the technical difficulty in isolating the retina alone, entire ocular globes were homogenized for assay. TBI significantly reduced VEGF, HGF, and IGFBP-6 levels in the eye, while MSC therapy markedly reversed these changes (except VEGF). (D) Western blot analysis of IGF-1R, VEGFR2, phosphorylated VEGFR2 (p-VEGFR2), and c-Met in R28 cell lysates. SI reduced receptor expression, while MSC coculture significantly restored it. Densitometric quantification is normalized to β-actin. (E) Western blot analysis of downstream signaling proteins in R28 cells. MSC coculture upregulated phosphorylated Akt (p-Akt), Sirtuin 1 (SirT1), and β-catenin in SI-injured cells. Quantified values are normalized to β-actin (p < 0.0001). Abbreviations: HGF, hepatocyte growth factor; IGFBP-4/6, insulin-like growth factor binding protein-4/6; VEGF, vascular endothelial growth factor; IGF-1R, insulin-like growth factor 1 receptor; VEGFR2, vascular endothelial growth factor receptor 2; p-Akt, phosphorylated Akt; SirT1, Sirtuin 1. Data are presented as mean ± SD. In vitro experiments (A, D, E) were performed in at least three independent replicates. In vivo data (B, C) are from n=8-9 (serum ELISA) or n = 6 (ocular tissues ELISA) animals per group. For original blot images, see the Supplementary Data file.
Article Snippet: IGFBP-6 , 1:200 , Bioss , BS-4064R , IF.
Techniques: Expressing, Ab Array, Enzyme-linked Immunosorbent Assay, Western Blot, Binding Assay, In Vitro, In Vivo