Salvianolic acid B promotes angiogenesis and inhibits cardiomyocyte apoptosis by regulating autophagy in myocardial ischemia

Figures

• 

  Fig. 1. The chemical structure of Sal B

• 

  Fig. 2. Sal B promoted the angiogenesis after glucose deprivation. A Sal B increased the angiogenesis in rats (4×, n = 4); B Sal B promoted the tubule formation in HUVECs (4×, n = 5); C Sal B facilitated the migration of HUVECs (4×, n = 5); D Sal B boosted the secretion of VEGF (n = 5); E The model of co-culture; F Sal B increased the expressions of VEGF and PDGF in H9c2 (n = 3); G Sal B increased the expressions of VEGF and PDGF in HUVECs (n = 3); Data are shown as the mean ± SD. *P < 0.05, **P < 0.01, compared to GD 24 h without Sal B group

• 

  Fig. 3. The deprivation of glucose aggravated the apoptosis of H9c2. A Effects of glucose deprivation at different time points on cell viability of H9c2 (n = 6); B Hoechst staining showed the apoptosis of H9c2 (20×, n = 3); C Flow cytometry revealed apoptosis of H9c2 (n = 3); D WB assayed the expressions of Bax and Bcl-2 (n = 3); Data are shown as the mean ± SD. *P < 0.05, **P < 0.01, compared to GD 0 h group

• 

  Fig. 4. Sal B alleviated the apoptosis of H9c2 induced by glucose deprivation. A Hoechst staining showed Sal B reduced the apoptosis of H9c2 (20×, n = 3); B WB showed Sal B regulated the expressions of Bax and Bcl-2 (n = 3); C Immunofluorescence staining showed Sal B reduced the level of ROS (20×, n = 3); D Sal B reduced the content of LDH, SOD and MDA (n = 6); E JC-1 staining showed Sal B elevated the mitochondrial membrane potential (20×, n = 3); Data are shown as the mean ± SD. *P < 0.05, **P < 0.01, compared to GD 24 h without Sal B group

• 

  Fig. 5. Sal B promoted the autophagy of H9c2 induced by glucose deprivation. A Electron microscopy showed Sal B promoted the formation of autophagosome (Scale bar = 0.2 μm, n = 3); B Immunofluorescence staining showed Sal B increased the expression of LC3 (20×, n = 3); C, D WB showed Sal B regulated the expressions of LC3, Beclin1, Atg5 and P62 (n = 3); Data are shown as the mean ± SD. *P < 0.05, **P < 0.01, compared to GD 24 h without Sal B group

• 

  Fig. 6. Sal B regulated the angiogenesis and apoptosis through Atg5. A The expression of Atg5 was inhibited by Atg5 siRNA (n = 3); B With the transfection of Atg5 siRNA, WB showed the effects of Sal B on regulating the expressions of LC3, Beclin1, Atg5 and P62 (n = 3); C Immunofluorescence staining showed the effect of Sal B on increasing the expression of LC3 was reduced by Atg5 siRNA (20×, n = 3); D The effect of Sal B on promoting the tubule formation in HUVECs was alleviated by Atg5 siRNA (4×, n = 5); E The effect of Sal B on increasing the expressions of VEGF and PDGF in H9c2 was alleviated by Atg5 siRNA (n = 3); F The effect of Sal B on increasing the expressions of VEGF and PDGF in HUVECs was alleviated by Atg5 siRNA (n = 3); G Hoechst staining showed the effect of Sal B on reducing the apoptosis of H9c2 was alleviated by Atg5 siRNA (20×, n = 3); H Immunofluorescence staining showed the effect of Sal B on reducing the level of ROS was alleviated by Atg5 siRNA (20×, n = 3); I JC-1 staining showed the effect of Sal B on elevating the mitochondrial membrane potential was reduced by Atg5 siRNA (20×, n = 3); J With the transfection of Atg5 siRNA, WB showed the effects of Sal B on…

• 

  Fig. 7. Sal B improved ISO induced myocardial ischemia in mice through Atg5. A Overall flow chart of animal experiment and changes in mice electrocardiogram. B Analysis of myocardial injury by HE staining (Scale bar = 50 μm, n = 5); C Analysis of myocardial fibrosis by Masson staining (Scale bar = 50 μm, n = 5); D The levels of LDH, AST, CK and Ca2+ were discovered using ELISA kits (n = 5); E The expressions of Bax and Bcl-2 in mice myocardial tissue were assayed by WB (n = 3); F The levels of NO, NOS, and eNOS were detected by ELISA kits (n = 5); G The angiogenesis pores on the surface of myocardial tissue with the use of scanning electron microscopy (Scale bar = 2 μm, n = 3); H ELISA kits were used to measure the levels of PDGF and VEGF in the serum (n = 5); I The expression of Atg5 in mice myocardial tissue was detected by immunohistochemistry (n = 3); J The expressions of VEGF, PDGF, and CD31 in mice myocardial tissue were detected by WB (n = 3); K The expressions of LC3 I, LC3 II, Beclin1 and P62 in mice myocardial tissue were detected by WB (n = 3); Data are shown as the mean ± SD. *P < 0.05, **P < 0.01, compared to ISO with control siRNA group