Figure 7.

AD-9308 improved mitochondrial functions and calcium homeostasis in streptozotocin (STZ)-induced diabetic mice and high-glucose and high-palmitate treated H9c2 cardiomyoblasts. (a) The activities of mitochondrial electron transfer chain complex were measured in cardiac tissue from STZ-induced diabetic mice treated with 0, 60, 180 mg/kg/day of AD-9308 by oral gavages and non-diabetic mice; (b) Mitochondrial respiration and (c) mitochondrial oxygen consumption rate in high-glucose and high-palmitate cultured H9c2 cells without or with AD-9308 treatment were measured. Data are presented as mean ± SEM (n = 3–6 per group). p-for-trend was used to test the linear trend. (d) Fluorescence spectra from Fura-2-AM loaded H9c2 cells were analyzed under high-glucose and high-palmitate medium-cultured condition (pretreated with 0, 5, and 10 μM AD-9308) and normal condition (control), and (e) represent Fura2-AM fluorescent images of free Ca²⁺ release from sacroendoplamic reticulum. (f) Immunoblottings of mitochondrial dynamic regulators, including long-form Optic atrophy 1 (L-Opa1), short-form Optic atrophy 1 (S-Opa1) and dynamin-related protein 1 (Drp1), and autophagy regulators, including Beclin1and Microtubule-associated proteins 1A/1B light chain 3A and 3B (LC3A/B) protein I and II, protein expressions and (g) representative immunohistochemical staining of LC3 were detected in cardiac tissue from STZ-induced diabetic mice treated with 0, 60, 180 mg/kg/day of AD-9308 by oral gavages and non-diabetic control mice.