Figure 8.

AT1R inhibition improved the imbalance of excessive mitochondrial fission and induced Rab9-dependent mitochondrial autophagy, leading to anti-vascular senescence. (A) Electron microscopic images and quantitative analysis of mitochondria and average mitochondrial areas in ApoE KO mice treated with Candesartan or the control. Scale bar = 1 μm. ^*P < 0.05 vs. ApoE KO mice with the control (n = 3 per group). (B) Immunoblots and quantitative analysis for Drp1, p-Drp1 (Ser616), p-Drp1 (Ser637), Mfn1, Mfn2, Opa1, and α-tublin in ApoE KO mice treated with Candesartan or the control. ^*P < 0.05 vs. ApoE KO mice with vehicle (n = 3 per group). (C) Relative mitochondrial H₂O₂ production in ApoE KO mice treated with Candesartan or the control. ^*P < 0.05 vs. ApoE KO mice with the control (n = 3 per group). (D) Relative mitochondrial ATP production in ApoE KO mice treated with Candesartan or the control. ^*P < 0.05 vs. ApoE KO mice with the control (ApoE KO mice with Candesartan; n = 5, ApoE KO mice with the control; n = 3). (E) Representative images and quantitative analysis of Masson's trichrome staining in ApoE KO mice treated with Candesartan or the control. Scale bar = 100 μm. ^*P < 0.05 vs. ApoE KO mice with the control (n = 3 per group). (F) Representative images and quantitative analysis of SA-β gal staining of arteries in ApoE KO mice treated with Candesartan or the control. ^*P < 0.05 vs. ApoE KO mice with the control vehicle (ApoE KO mice with Candesartan; n = 3, ApoE KO mice with the control; n = 4). (G) Immunoblots and quantitative analysis for p53, p21, and β-actin in ApoE KO mice with Candesartan or the control. ^*P < 0.05 vs. ApoE KO mice with the control (n = 3 per group). (H) Electron microscopic images of mitochondrial autophagy in ApoE KO mice treated with Candesartan or the control. Scale bar = 1 μm. ^**P < 0.01 vs. ApoE KO mice with the control (n = 14 per group). (I) Representative images of TOMM20 (green) and LAMP2 (red) immunohistochemistry in ApoE KO mice treated with Candesartan or the control. Scale bar = 10 μm. ^*P < 0.05 vs. ApoE KO mice with the control (n = 3 per group). (J) Representative images and quantitative analysis of LAMP2 (green) and Rab9 (red) immunohistochemistry in the ApoE KO mice treated with Candesartan or the control. Scale bar = 10 μm. ^*P < 0.05 vs. ApoE KO mice with the control (n = 3 per group). All Data are presented as the mean ± SEM. Statistical analysis were conducted by Wilcoxon rank sum test (A-G,I,J) and Student's t-test (H).