Empagliflozin Reverses Oxidized LDL-Induced RECK Suppression, Cardiotrophin-1 Expression, MMP Activation, and Human Aortic Smooth Muscle Cell Proliferation and Migration

Figure 7

Empagliflozin inhibits OxLDL and CT-1-induced SMC proliferation and migration. (a) Empagliflozin inhibits OxLDL-induced miR-30b expression. Quiescent SMCs were treated with empagliflozin for 15 min followed by the addition of OxLDL or nLDL for 2 hr and analyzed for miR-30b expression by RT-qPCR using a TaqMan® probe. (b, c) Empagliflozin reverses OxLDL-induced RECK suppression. Quiescent SMCs were treated with empagliflozin as in (a), were exposed OxLDL or nLDL for 6 hr and analyzed for RECK expression by western blotting. Semiquantification of the intensity of immunoreactive bands by densitometry is shown in panel (c). (d, e) Empagliflozin inhibits OxLDL- or CT-1-induced MMP2 (d) and MMP9 (e) activity. SMCs made quiescent in medium supplemented with ITS-G 1x and no FBS were exposed to empagliflozin as in (a), followed by OxLDL (black) or CT-1 (red) for 18 hr and analyzed for MMP2 and MMP9 activity using SensoLyte® 520 fluorimetric assay. (f, g) Empagliflozin inhibits OxLDL- or CT-1-induced SMC proliferation and migration. Quiescent SMCs exposed to empagliflozin were treated with OxLDL (black) or CT-1 (red) for 48 hr (f) or 18 hr (g) and then analyzed for proliferation. (h) Empagliflozin reverses OxLDL-induced suppression of the mRNA expression of SMC markers αSMA and MYH11 and the proinflammatory markers ICAM1, Galectin 3, and Olr1 as analyzed by RT-qPCR. (a) versus nLDL, ^† versus OxLDL (n = 5); (c) versus nLDL, ^† versus OxLDL (n = 3); (d–g) versus nLDL, ^† versus OxLDL or CT-1 (n = 4 or 5), (h) versus nLDL, ^† versus OxLDL (n = 5).

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