Research Paper Volume 12, Issue 2 pp 1141—1158

MiR-29b-3p promotes particulate matter-induced inflammatory responses by regulating the C1QTNF6/AMPK pathway

Figure 2. MiR-29b-3p promoted PM-induced inflammatory responses via repressing AMPK pathway activation. (A) HBECs were transfected with miR-29b-3p mimic (29b-m) or negative control mimic (NC-m), and then treated with or without 300 μg/cm3 PM for 24 h. Real-time PCR analysis of IL-1β, IL-6, and IL-8 expression in HBECs transfected with 29b-m or NC-m prior to PM exposure. Values represent mean ± SEM; *, P<0.05, compared with the NC-m + PM group; #, P<0.05, compared with the NC-m group; n=3. (B) HBECs were transfected with miR-29b-3p inhibitor (29b-i), or negative control inhibitor (NC-i), and then treated with or without 300 μg/cm3 PM for 24 h. Real-time PCR analysis of IL-1β, IL-6, and IL-8 expression in HBECs transfected with 29b-i or NC-i prior to PM exposure. Values represent mean ± SEM; *, P<0.05, compared with the NC-i + PM group; #, P<0.05, compared with the NC-i group; n=3. (C) Western blot analysis of AMPK signaling pathway activation in HBECs transfected with 29b-m or NC-m prior to PM exposure. The optical densities of protein bands were shown in (D). Values represent mean ± SEM; *, P<0.05, compared with the NC-m + PM group; n=3. (E) The AMPK siRNAs were transfected into HBECs 24h prior to PM exposure, respectively and the optimum AMPK siRNA was selected using real-time PCR. Values represent mean ± SEM; *, P<0.05, compared with the NC siRNA group; n=3. (F) Real-time PCR analysis of IL-1β, IL-6, and IL-8 expression in HBECs transfected with AMPK siRNA or negative control siRNA prior to PM exposure. Values represent mean ± SEM; *, P<0.05, compared with the NC siRNA + PM group; #, P<0.05, compared with the NC siRNA group; n=3. HBECs, human bronchial epithelial cells; PM, particulate matter.