Resveratrol Inhibits ROS-Promoted Activation and Glycolysis of Pancreatic Stellate Cells via Suppression of miR-21

Figure 5

miR-21 is essential for RSV-mediated responses of PSCs. (a) PSCs were transfected with miR-21 mimics (20 nM) or scrambled mimics (20 nM) for 24 h, and RT-PCR analysis was performed to detect miR-21 expression. (b–d) PSCs were treated with 50 μM RSV for 24 h and then transfected with miR-21 mimics (20 nM) or scrambled mimics (20 nM) prior to the 24 h incubation with H₂O₂ at 50 μM. PSCs from indicated groups were extracted to detect α-SMA, Glut1, HK2, PKM2, LDHA, and PTEN levels by Western blot, and the CM were collected to measure lactate production. Lactate production was normalized by the concentration of protein in each group. (e, f) Immunofluorescence staining of α-SMA was performed. The red signal represents α-SMA staining, and nuclear DNA staining by DAPI is shown in blue (magnification, ×200; scale bar: 50 μm). (g, h) PSCs were treated in groups as indicated, and migratory ability was determined by wound healing assays. The relative distance moved by the leading edge marked by black lines was measured 24 h after wounding with a sterile pipette tip (magnification, ×100; scale bar: 100 μm). (i, j) PSCs were treated in groups as indicated, and invasive ability was determined by Matrigel invasion assays (magnification, ×200; scale bar: 50 μm). Column: mean; bar: SD; compared with the control group; compared with the H₂O₂ group; compared with the RSV + scramble + H₂O₂ group.

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