Article Figures & Data
Figures
- Fig. 1.
Experimental design and protocol. The schematic diagram shows the sequence and timing of various experimental procedures, including the drug treatment, OGTT, insulin tolerance test (ITT), harvest of tissue, global I/R in Langendorff mode, histology analysis, as well as plasma and cardiac tissue biochemical measurements.
- Fig. 2.
Effect of TAD, HCQ, and combination treatment on postischemic infarct size in db/db mice. Top: Representative images of transverse sections of TTC-stained hearts collected after 1-week respective drug treatments and ex vivo global I/R. Bottom: Bar diagram showing myocardial infarct size presented as % of risk area (mean ± S.E.M., n = 6 for CTRL and n = 5 for TAD, HCQ, and TAD + HCQ). *P < 0.05 versus CTRL group.
- Fig. 3.
Effect of TAD, HCQ, and combination treatment on cardiac function. Post-I/R ventricular developed force (A), rate-force product (B), heart rate (C), and coronary flow rate (D) are presented as % of preischemia baseline (mean ± S.E.M.; n = 6 for CTRL and n = 5 for TAD, HCQ, and TAD + HCQ). No statistical significance was observed among the four treatment groups in any of the cardiac function indices.
- Fig. 4.
Changes in blood glucose, plasma insulin, IGF-1 levels, and lipid profile after treatment with TAD and HCQ. (A, B) Plasma insulin and IGF-1 levels measured with ELISAs. (C) Blood glucose levels measured before and after the 1-week drug treatment, respectively (mean ± S.E.M.; n = 13 for CTRL and HCQ, n = 15 for TAD, and n = 12 for TAD + HCQ). (D–H) Plasma and cardiac levels of free fatty acids, triglycerides, and total cholesterol measured using enzymatic assays (mean ± S.E.M., n = 8 for TAD, n = 9 for HCQ, and n = 12 for CTRL and TAD + HCQ). *P < 0.05; ***P < 0.001 versus CTRL group.
- Fig. 5.
Insulin tolerance test (ITT) and OGTT after treatment with TAD, HCQ, and a combination of TAD + HCQ. Animals were fasted overnight before the OGTT. Glucose (2 mg/kg) was administered via oral gavage, and blood samples were taken from the tails. (A) Glucose levels; (B) area under the curve; (C) insulin levels; and (D) area under the curve. The ITT was performed after the animals were fasted for 6 hours. Insulin (0.9 IU/kg regular human) was given i.p.; (E) blood glucose levels; (F) area under the curve; (G) insulin response curve is presented as percentage to baseline glucose. Data are mean ± S.E.M. (n = 5/group,). *P < 0.05 versus CTRL group.
- Fig. 6.
Effect of TAD, HCQ, or combination treatment on pancreatic islets. (A) Representative pictures of immunofluorescent-stained paraffin sections of pancreata. Goat anti-insulin antibody was used to detect insulin inside islets, and a secondary antibody conjugated with fluorescein isothiocyanate was used. (B) Representative pictures of H&E-stained pancreas, paraffin-fixed sections, with further magnified representative images of the pancreatic islets at lower-right corners. (C) Insulin-positive β-cell area versus pancreas area (n = 5/group). (D) Bar diagram showing pancreatic islet number per mm2 pancreas area in all treatment groups (n = 8 for TAD; n = 9 for CTRL, HCQ, and TAD + HCQ). (E) Bar diagram showing the percentage of pancreas mass versus body weight (n = 5/group). Data are mean ± S.E.M. *P < 0.05; **P < 0.01 versus CTRL.
- Fig. 7.
Effect of TAD, HCQ, or combination treatment on mTOR activation after I/R. (A) Representative Western blot images; (B) bar diagram showing quantitative analysis of cardiac p-AktThr308/Akt; (C) p-mTOR/mTOR; (D) expression of Raptor; (E) p-S6/S6; (F) Rictor; and (G) p-AktSer473/Akt after I/R. Data are presented as mean ± S.E.M. (n = 4/group). *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001 versus CTRL group.
- Fig. 8.
Effect of TAD, HCQ, and combination treatment on myocardial ATP production. ATP levels are normalized with respective protein concentration for each sample. Data are presented as mean ± S.E.M. (n = 4/group). **P < 0.01 versus CTRL group.
