Dysregulation in microRNA Expression Is Associated with Alterations in Immune Functions in Combat Veterans with Post-Traumatic Stress Disorder
Figure 5
Dysregulation in miRNA expression profile in PBMC from PTSD patients and controls.
A–F: Total RNA from PBMC of patients with PTSD and normal controls were used in the analysis of miRNA expression by Affymetrix miRNA array hybridization. Comparison of miRNA expression profiles between individual controls and PTSD patients was shown by the heat map (A) and plot of the principal component analysis accounting for 60.1% variability (B). Seven up-regulated miRNAs in PBMC from PTSD patients were compared with those from controls (C). 64 down-regulated miRNA molecules (>2.5 fold change, Table 1) in PBMC from PTSD patients were detected (D). Total RNA samples were also used in the confirmation of miRNA down-regulation (miR-125a and miR-181c) in PBMC from PTSD patients by real-time PCR (E). Wilcoxon rank sum test was used to compare the difference in miRNA expression in PBMC between controls and PTSD patients. F–G: Role of hsa-miR-125a in the regulation of IFN-γ. Complementary sequences between the seed sequence of miR-125a and 3′-untranslated region (3′UTR) of IFN-γ gene were compared (F). In silico studies were used to determine the complementary sequences between the seed sequence of miR-125a and 3′UTR of IFN-γ gene. The inhibitory effect of miR-125a on IFN-γ production in PBMC was determined (G). Hsa-miR-125a precursor (pre-miR125a) and pre-scramble control (scrambled premiR) were introduced into PBMC by electroporation. After PHA stimulation for two days, IFN-γ release from PBMC was determined in the culture supernatant by ELISA. Wilcoxon rank sum test was used to analyze the inhibitory effects of miR-125a on IFN-γ production in PBMC.