Figure 1
Over-expression of mir-21 inhibited apoptosis and promoted collagen synthesis by activating the ERK/NF-κB /NLRP3 inflammasome pathway via targeting Spry1 in lung fibroblasts. Primary lung fibroblasts were transfected with control miRNA precursors (pre-NC) or mir-21 precursors (pre-21) for 48 h and then treated with U0126 (10−5 mol/L) or BAY 117082 (10−5 mol/L). (A) The expression of mir-21 in lung fibroblasts was determined by qRT-PCR. (B) The protein levels of Spry1, p-ERK, and NF-κB were analyzed by western blot. (C) The protein levels of NLRP3, pro-caspase-1 (pro-casp-1), cleaved caspase-1 (cleaved casp-1), Pro-IL-1β, active IL-1β, and a-collagen I were analyzed by western blot. (D) Dual immunofluorescence of NLRP3 (red) and caspase-1 (green). Nuclei were stained with DAPI (blue). The yellow Fluorescence units were measured, which represent the co-location of NLRP3 and casp-1. The images are representative of 3 separate experiments. (E) Apoptosis of lung fibroblasts was evaluated by flow cytometry. Scale bar = 20 μm. Data are the means ± SD from 3 independent experiments. *P < 0.05 versus NC; #P < 0.05 versus pre-21.
