Fig 2: mRNA expression variability of carotid endarterectomy plaques by lesion severity and diabetic status (A–C) No significant difference in carotid artery plaque expression between the Min (n = 18) and Max (n = 18) segments of ppara, acox1, and cpt1a mRNA expression. (D–F) Combined Min and Max analysis of patients with diabetes (n = 18) demonstrate significantly higher mRNA expression of ppara (p < 0.05), and acox1 (p < 0.001) compared to patients without diabetes. We did not observe a difference between cpt1a mRNA expression between patients with or without diabetes. (G–I) When distinguishing both disease severity and diabetic status, we found significant increase in ppara mRNA expression in patients with diabetes (n = 9) compared to patients without diabetes (n = 9). We found no significant differences in acox1 or cpt1a expression. DM, Diabetic. ND, Non-diabetic. Min, Minimally diseased. Max, Maximally diseased. Data are presented as mean ± SD, and each point represents one patient sample. *p < 0.05 and ***p < 0.001.

Fig 3: mRNA expression variability of lower extremity segments by lesion severity and diabetic status: (A–C) In Lower extremity samples, we observed no significant difference in ppara and cpt1a mRNA expression between Min (n = 18) and Max (n = 18) diseased segments. However, we found Min-diseased samples had non-significantly lower expression of acox1 (p = 0.051, n = 18) compared to Max-diseased. (D–F) We found that patients with diabetes (n = 20) demonstrated significantly lower ppara (p < 0.05), acox1 (p < 0.001) and cpt1a (p < 0.0001) mRNA expression compared to patients without diabetes (n = 16). (G–I) When categorizing segments between diseased and diabetic status, we observed no significant variability of ppara. However, we found a significant decrease of acox1 (p < 0.01) and cpt1a (p < 0.01) in patients with diabetes (n = 10) compared to patients without diabetes (n = 8) from Min-diseased tissues. DM, Diabetic. ND, Non-diabetic. Min, minimally diseased. Max, maximally diseased. Data are presented as Mean ± SD and, each point represents one patient sample. *p < 0.05, **p < 0.01, and ***p < 0.001.

Fig 4: Dapagliflozin alleviates hepatic lipid accumulation in ZDF rats and PA-stimulated LO2 and HepG2 cells. (A, B) Liver sections were stained with H&E (Scale bars: 50 μm)and Oil Red O (Scale bars: 20 μm). (C) Liver/body weight ratio was calculated by liver weight (g)/body weight (g). (D) The mRNA expression levels of hepatic lipogenic genes ACC1, SREBP-1c, and fatty acid oxidation genes CPT1 and ACOX1 were examined by real-time q-PCR (E) CCK-8 assays indicate the effect of dapagliflozin (10, 20, 40, 80, and 100 μmol/L) on the viability of HepG2 cells and LO2 cells. (F) HepG2 cells and LO2 cells were stained with 1 μg/ml BODIPY 493/503 and fluorescence images were captured by a confocal microscope. (G) LO2 and HepG2 cells were stained with Oil Red O. Scale bars: 20 μm. Data are expressed as the means ± SEM from three independent experiments. *p < 0.05, **p < 0.01, and ***p < 0.001.

Fig 5: The luciferase assay of miR-15a targeting the 3′UTR of ACAA1, ACOX1 and SCP2 in DF1 cells. (A–D) denote the expression levels of miR-15a, ACAA1, ACOX1 and SCP2 in DF1 cells transfected with miR-15a mimics, respectively. (E) Results from tests in which ACAA1 mRNA 3′ UTR or its mutation in a dual-luciferase vector were co-transfected with miR-15a mimics/NC. (F) Results from tests in which ACOX1 mRNA 3′ UTR or its mutation in a dual-luciferase vector were co-transfected with miR-15a mimics/NC. (G) Results from tests in which SCP2 mRNA 3′ UTR or its mutation in a dual-luciferase vector were co-transfected with miR-15a mimics/NC. The four colored bars in (E–G) indicate co-transfection tests with four combinations, respectively. Data are expressed as mean ± SEM (n = 3); * p < 0.05; ** p < 0.01; ns, not significant. NC, negative control; mimics, miR-15a mimics.