Fig. 4

EXO-Hep improved mitochondrial function by inhibiting Drp1 / Fis1 mediated mitochondrial fission in type A1 astrocytes and enhancing the secretion of healthy astrocytic mitochondria. A The protocol for obtaining purified mitochondria in the culture medium. B A1 astrocytic activation was examined by detecting the expression of C3 (type A1 astrocyte marker) in different groups. Data represent means ± SD (n = 3), **P < 0.01, ***P < 0.001. C Western blotting assays on the expressions of Drp1 and Fis1 in intracellular mitochondria of type A1 astrocytes treated with Hep (A1-AS (Hep)), EXO (A1-AS (EXO)) and EXO-Hep (A1-AS (EXO-Hep)). Data represent means ± SD (n = 3), **P < 0.01, ***P < 0.001. D Extracellular ATP levels, mitochondrial ROS, and mitochondrial membrane potential, were determined in released Mito/AS, Mito/A1-AS, Mito/A1-AS(Hep), Mito/A1-AS(EXO) and Mito/A1-AS(EXO-Hep). The relative ratio of extracellular ATP was by calculating the ratio of level of ATP in groups to level of ATP in released Mito/AS group. Data represent means ± SD (n = 3), **P < 0.01, ***P < 0.001. E Integrity of the released Mito/AS, Mito/A1-AS, Mito/A1-AS (Hep), Mito/A1-AS (EXO) and Mito/A1-AS (EXO-Hep) was determined using western blot for detecting markers of outer-membrane proteins (TOM20) and cytochrome c as an inter-membrane space protein in the released mitochondria, the mitochondrial reference VDAC was used as a reference. Data represent means ± SD (n = 3),*P < 0.05