MicroRNA-1224-5p Aggravates Sepsis-Related Acute Lung Injury in Mice

Figure 7

miR-1224-5p directly targets PPAR-γ to regulate AMPKα activation. (a) The predicted miR-1224-5p-binding site within the PPAR-γ 3-UTR. (b) The PPAR-γ 3-UTR luciferase reporter plasmid was transfected to the HEK293T cells for 48 h with or without the miR-1224-5p agomir treatment. Next, cells were lysed and the luciferase activities were measured. (c, d) Mice were intravenously injected with the agomir (30 mg/kg/day), antagomir (80 mg/kg/day), or matched controls for 3 consecutive days and then exposed to LPS (5 mg/kg) for 12 h. PPAR-γ protein and mRNA levels in lungs were detected. (e, f) To inhibit PPAR-γ, mice were pretreated with GW9662 (0.35 mg/kg/day in drinking water) for 10 consecutive days, and then, phosphorylated and total AMPKα in lungs were detected. (g) ROS levels in lungs were measured. (h) The levels of IL-6 and TNF-α in murine lungs. (i) Lung wet to dry ratio. (j) LDH activities in lungs. (k) Tidal volume was detected. (l) Arterial blood gas analysis of PaO₂ and PaCO₂. Data represent the ( per group). versus the matched group.

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