circ-FoxO3 is upregulated and autophagy is activated in the BBB after cerebral I/R injury
(A and B) Images of circ-FoxO3 and CD31 and their quantification (integrated optical density) in brain tissues at the site of intracerebral hemorrhage from patients with HT and the peri-infarct area of MCAO/R mice. Ctrl group: brain tissues from non-stroke patients (six females and six males). HT group: patients with hemorrhagic transformation, six females and eight males. Sham group: male mice without ligation of the arteries and insertion of nylon monofilaments, n = 6. MCAO/R group: male mice under MCAO and reperfusion for 2 h, respectively, n = 6. White arrowheads indicate the co-localization of circ-FoxO3 and CD31. Scale bars, 50 μm. (C) The ultra-structures of the BBB in human. The cross-section of the BBB from patients with glioma and patients with HT were imaged in I and II, respectively. The autophagosome-like vacuoles were enlarged in III to VI. Scale bar, 500 nm. (D) Quantitative number of autophagosome-like vacuoles in the BMECs from patients with HT. n (Ctrl group): three females and three males. n (HT group): three females and four males. (E) Ultra-structure of BBB in mice. The cross-section of BBB from sham and MCAO/R mice was presented in I and II, respectively. Cell-cell junctions marked # in I and II was enlarged in III and IV, respectively. White dotted line: cell-cell junctions. Arrowheads indicate desmosomes. Scale bar, 500 nm. The autophagosome-like vacuoles enlarged into V to VIII. Scale bar, 500 nm. (F) Quantitative number of autophagosome-like vacuoles in the BMECs from male MCAO/R mice. n = 6. ECs, endothelial cells. Black arrowheads, autophagosome-like vacuoles; L, lumen; #, cell-cell junction; P, pericyte; Asp, astrocyte processes; MG, microglia; MS, medullary sheath; M, mitochondria; N, nucleus; AP, autophagosome; AL, autolysosome; MCAO/R, middle cerebral artery occlusion for 2 h followed by reperfusion for 2 h. p values indicate a two-tailed unpaired Student's t test. Data are provided as the mean ± SEM. ∗p < 0.05, ∗∗p < 0.01.