Tanshinone IIA Inhibits β-Catenin Nuclear Translocation and IGF-2R Activation via Estrogen Receptors to Suppress Angiotensin II-Induced H9c2 Cardiomyoblast Cell Apoptosis

Figure 1. TSN suppressed the AngII-induced activation of β-catenin and IGF-2pathways and increased the expression of endogenous ERs and survival pathway in H9c2 cardiomyoblasts in a dose-dependent manner; all the experiments were repeated at least three times. (A) Twenty-four hours after seeding the H9c2 cells in 10-cm culture plates, different doses of AngII were added. Western blot analysis was conducted 24 h after the administration of AngII. (B) The difference is statistically significant. (C) Twenty-four hours after seeding the H9c2 cells in 10-cm culture plates, 10-8 M AngII was added, followed by different doses of TSN. Western blot analysis was conducted 24 h after the administration of TSN. (D) Difference is statistically significant. Data are expressed as the fold change relative to the control and presented as the mean ± SD, representing the results of three independent experiments (n = 3, *p < 0.05 was considered significant).

Figure 2. TSN reduced AngII-induced β-catenin expression and nuclear localization through ERs; all the experiments were repeated at least three times. (A) Twenty-four hours after seeding the H9c2 cells in 10-cm culture plates,10-8M AngII was added, followed by 40μM TSN and 10-6 M ICI. Western blot analysis was conducted 24 h after the administration of ICI. (B) fold change were calculated and are shown. (C) Twenty-four hours after seeding the H9c2 cells in 10-cm culture plates, 10-8 M AngII was added, followed by 40μMTSN and 10-6 M ICI. The nuclear and cytoplasmic fractions were separated 24 h after the administration of ICI. (D) Difference is statistically significant. (E) Twenty-four hours after seeding the H9c2 cells in 10-cm culture plates, 10-8 M AngII was added, followed by 40μM TSN and 10-6 M ICI. Confocal microscopy was conducted 24 h after the administration of ICI. Data are expressed as the fold change relative to the control and presented as the mean ± SD, representing the results of three independent experiments (n = 3, *p < 0.05 was considered significant).

Figure 3. TSN reduced AngII-induced apoptosis and promoted survival via estrogen receptors in H9c2 cardiocytes. (A) Twenty-four hours after seeding the H9c2 cells in 10-cm culture plates, 10-8 M AngII was added, followed by 40μM TSN and 10-6 M ICI. Western blot analysis was conducted 24 h after the administration of ICI. (B) Fold change were calculated and are shown. (C) Twenty-four hours after seeding the H9c2 cells in 10-cm culture plates, 10-8M AngII was added, followed by 40μM TSN and 10-6 M ICI. TUNEL staining was conducted 24 h after the administration of ICI (400X magnification). Data are expressed as the fold change relative to the control and presented as the mean ± SD, representing the results of three independent experiments (n = 3, *p < 0.05 was considered significant).

Figure 4. TSN reduced AngII-induced fibrosis via ERs in H9c2 cardiomyocytes. (A) Twenty-four hours after seeding the H9c2 cells in 10-cm culture plates, 10-8M AngII was added, followed by 40μM TSN and 10-6 M ICI. Western blot analysis was conducted 24 h after the administration of ICI. (B) Fold change were calculated and are shown.