Figure 7.

MicroRNA (miR)-378a-3p induces metabolic reprogramming and promotes smooth muscle cells (SMC) differentiation. A, The oxygen consumption rate (OCR) of perivascular adipose tissue-derived mesenchymal stem cells (PV-ADSCs) cultured in miR-378a-3p mimics or control for 2 d (n=3). B, Live PV-ADSCs treated with miR-378a-3p mimic for 2 d with or without TGF (transforming growth factor)-β1 were stained tetramethylrhodamine, methyl ester, perchlorate (TMRM) and analyzed for mean fluorescence intensity (MFI) with flow cytometry (n=3). C and D, Cellular metabolite abundance in PV-ADSCs treated with miR-378a-3p mimics for 1 d determined with gas chromatography-mass spectrometry (GC-MS) system (n=3). Heatmap of metabolites in tricarboxylic acid (TCA) cycle (C) or lipid metabolism (D). E, SMC marker mRNAs were induced by miR-378a-3p mimics after 2 d (n=3). F, Representative immunofluorescent staining (n=3) showed the upregulation of SMC markers by miR-378a-3p mimic treatment. G, Transfection of PV-ADSCs with miR-378a-3p inhibitor inhibited the level of SMC markers as shown by quantitative polymerase chain reaction (qPCR). The level of Crat was derepressed by miR-378a-3p inhibitor transfection (n=3). H, OCR and extracellular acidification rate (ECAR) were detected with Seahorse Mito stress tests in wild-type (WT) and miR-378a knockout (KO) ADSCs (n=3). I, WT and miR-378a KO ADSCs were treated with or without TGF-β1 for 2 d. Immunofluorescent staining showed the protein level of CNN1 (calponin; n=1). Scale bar, 100 µm. Data are mean±SD. *P<0.05 and **P<0.01. Anti-A/Rot indicates anti-mycin A/Rotenone; Crat, carnitine acetyltransferase; FCCP, carbonyl cyanide-4-phenylhydrazone; inh 378, miR-378a-3p inhibitor; inh ctrl, inhibitor control; mim 378, miR-378a-3p mimics; mim ctrl, mimics control; and SRC, spare respiratory capacity.