Fig. 6: Mitochondrial Dysfunction in SH-SY5Y Cells by TSPO Overexpression Vector (pLV-TSPO) and PK11195 and Schematic Diagram. | Translational Psychiatry

Fig. 6: Mitochondrial Dysfunction in SH-SY5Y Cells by TSPO Overexpression Vector (pLV-TSPO) and PK11195 and Schematic Diagram.

From: Critical role of translocator protein (TSPO) in neuronal mitochondrial dysfunction and mental stress-exacerbated ischemic injury following stroke

Fig. 6

A CCCP induced mitochondrial membrane potential (MMP) disruption at doses of 25, 50, and 100 μM, and CCCP worsened OGD/R damage at 6 and 24 h after reoxygenation at doses of 25, 50, and 100 μM. B mito-TEMPO alleviated CCCP increased MMP at doses of 25 and 50 μM, with the 25 μM dose yielding the highest viability, and mito-TEMPO decreased OGD/R induced neuron damage at 6 and 24 h after reoxygenation at doses of 25 and 50 μM, with the 25 μM dose yielding the highest viability. C TSPO overexpression vector (pLV-TSPO) was constructed and transfected effectively into SH-SY5Y cell lines. D TSPO overexpression vector increased the mRNA (p < 0.001) and protein expression (p < 0.01) in SH-SY5Y cells. E TSPO decreased mPTP opening (p < 0.001), and increased intracellular Ca2+ by Fluo-4 staining (p < 0.01), the ratio of green and red fluorescence in JC1 staining (p < 0.01) and mitochondrial ROS by mito-SOX staining (p < 0.001). F PK11195 increased mPTP opening (p < 0.001), and decreased the intracellular Ca2+ (p < 0.01), the ratio of green and red fluorescence in JC1 staining (p < 0.01), and mito-ROS (p < 0.01). Data are presented as mean ± SEM. G This schematic diagram illustrates the molecular cascade triggered by mental stress that exacerbates ischemic stroke injury. Under mental stress conditions, expression of translocator protein (TSPO) is upregulated in the mitochondrial membrane. Increased TSPO interacts with ACBD3, activating the protein kinase A (PKA) signaling pathway, which subsequently phosphorylates the voltage-dependent anion channel (VDAC). This phosphorylation event disrupts mitochondrial function, leading to neuronal calcium/calmodulin-dependent protein kinase (CaMK) activation, increased intracellular calcium, oxidative stress, and neuronal injury. Collectively, this TSPO/ACBD3/PKA/VDAC1 phosphorylation/CaMK signaling pathway contributes significantly to mitochondrial dysfunction and is a critical mechanism by which mental stress aggravates ischemic brain damage.

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