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

Figure 6

OxLDL suppresses RECK expression via NF-κB-dependent miR-30b induction. (a) OxLDL upregulates miR-30b-5p (miR-30b) expression. Quiescent SMCs exposed to OxLDL or nLDL for 2 hr were analyzed for miR-30b expression by RT-qPCR using a TaqMan® probe. (b) OxLDL induces miR-30b expression in part via NF-κB. SMCs transduced with lentiviral NF-κB p65 or eGFP shRNA were made quiescent and then exposed to OxLDL or nLDL for 2 hr. miR-30b expression was analyzed by RT-qPCR. (c) miR-30b antagomir reverses OxLDL-induced RECK suppression. RECK expression was analyzed by western blotting, and the densitometric data is shown on the right. (d) miR-30b-mimic inhibits RECK expression. SMCs transfected with miR-30b-mimic or corresponding NC were analyzed for RECK expression by western blotting. Densitometric results are shown on the right. (e, f), miR-30b-antagomir attenuates OxLDL-induced SMC proliferation and migration. SMCs transfected with miR-30b-antagomir or corresponding NC were made quiescent and exposed to OxLDL or nLDL for 48 hr (e) or 18 hr (f) and analyzed for proliferation and migration. The inset in (f) shows representative images of Matrigel™ transwell invasion. (a, b) versus nLDL, ^† versus OxLDL or OxLDL + eGFP (n = 6); (c, d) versus nLDL, ^† versus OxLDL or OxLDL + NC (n = 3); (e, f) versus nLDL, ^† at least 0.05 versus OxLDL or OxLDL + NC (n = 5).

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