Description

Dr. Charles E. Norton from the University of Missouri will discuss his paper "Endothelial cells promote smooth muscle cell resilience to H2O 2-induced cell death in mouse cerebral arteries." Here is a link to the article: https://pubmed.ncbi.nlm.nih.gov/35380737/Authors: Charles E. Norton, Rebecca L.Shaw, Ron Mittler, Steven S. Segal 
Aim: Brain injury produces reactive oxygen species (ROS). However, little is known of how acute oxidative stress affects cell survival in the cerebral vascular supply. We hypothesized that endothelial cells (ECs) are more resilient to H₂ O₂ and protect vascular smooth muscle cells (SMCs) during acute oxidative stress.
Methods: Mouse posterior cerebral arteries (PCAs; diameter, ~80 µm) were exposed to H₂ O₂ (200 µM, 50 min, 37°C). Nuclear staining identified dead and live cells of intact and endothelium-disrupted vessels. SMC [Ca²⁺ ]_i was assessed with Fura-2 fluorescence, and superoxide production was assessed by dihydroethidium and MitoSOX fluorescence.
Results: In response to H₂ O₂ : SMC death (21%) exceeded EC death (5%) and increased following endothelial disruption (to 48%) with a corresponding increase in SMC Ca²⁺ entry through transient receptor potential (TRP) channels. Whereas pharmacological inhibition of TRPV4 channels prevented SMC death and reduced Ca²⁺ entry for intact vessels, both remained elevated following endothelial disruption. In contrast, pharmacological inhibition or genetic deletion of TRPC3 prevented SMC death and attenuated Ca²⁺ entry for both intact and endothelium-disrupted vessels. Inhibiting gap junctions increased EC death (to 22%) while SMC death and [Ca²⁺ ]_i responses were attenuated by inhibiting nitric oxide synthesis or scavenging superoxide/peroxynitrite. Inhibiting NADPH oxidases also prevented SMC Ca²⁺ entry and death. H₂ O₂ increased mitochondrial ROS production while scavenging mitochondria-derived superoxide prevented SMC death but not Ca²⁺ entry.
Conclusions: During acute exposure of cerebral arteries to acute oxidative stress, ECs are more resilient than SMCs and the endothelium may protect SMCs by reducing Ca²⁺ entry through TRPC3 channels.