Parallel-plate Chamber System Research Articles

Low shear stress inhibits endothelial mitophagy via caveolin-1/miR-7-5p/SQSTM1 signaling pathway

Background and aimsMitophagy plays a crucial role in mitochondrial homeostasis and is closely associated with endothelial function. However, the mechanism underlying low blood flow shear stress (SS), detrimental cellular stress, regulating endothelial mitophagy is unclear. This study aimed to investigate whether low SS inhibits endothelial mitophagy via caveolin-1 (Cav-1)/miR-7-5p/Sequestosome 1 (SQSTM1) signaling pathway. MethodsLow SS in vivo modeling was induced using a perivascular SS modifier implanted in the carotid artery of mice. In vitro modeling, low and physiological SS (4 and 15 dyn/cm2, respectively) were exerted on human aortic endothelial cells using a parallel plate chamber system. ResultsCompared with physiological SS, low SS significantly inhibited endothelial mitophagy shown by down-regulation of SQSTM1, PINK1, Parkin, and LC 3II expressions. Deficient mitophagy deteriorated mitochondrial dynamics shown by up-regulation of Mfn1 and Fis1 expression and led to decreases in mitochondrial membrane potential. Cav-1 plays a bridging role in the process of low SS inhibiting mitophagy. The up-regulated miR-7-5p expression induced by low SS was suppressed after Cav-1 was silenced. The results of dual-luciferase reporter assays showed that miR-7-5p targeted inhibiting the SQSTM1 gene. Oxidative stress reaction shown by the elevation of reactive oxygen species and O2●— and endothelial dysfunction by the decrease in nitric oxide and the increase in macrophage chemoattractant protein-1 were associated with the low SS inhibiting endothelial mitophagy process. ConclusionsCav-1/miR-7-5p/SQSTM1 signaling pathway was the mechanism underlying low SS inhibiting endothelial mitophagy that involves mitochondrial homeostasis impairment and endothelial dysfunction.

Transport and Retention of Free-Living Amoeba Spores in Porous Media: Effects of Operational Parameters and Extracellular Polymeric Substances.

Amoebas are protists that are widespread in water and soil environments. Some species are pathogenic, inducing potentially lethal effects on humans, making them a major threat to public health. Nonpathogenic amoebas are also of concern because they have the potential to carry a mini-microbiome of bacteria, either transiently or via more long-term stable transport. Due to their resistance to disinfection processes, the physical removal of amoeba by filtration is necessary to prevent their propagation throughout drinking water distribution networks and occurrence in tap water. In this study, a model amoeba species Dictyostelium discoideum was used to study the transport and retention behavior of amoeba spores in porous media. The key factors affecting the transport behavior of amoeba spores in fully saturated media were comprehensively evaluated, with experiments performed using a quartz crystal microbalance with dissipation monitoring (QCM-D) and parallel plate chamber system. The effects of ionic strength (IS) on the deposition of spores were found to be in contrast to the predicted Derjaguin-Landau-Verwey-Overbeek (DLVO) theory that more deposition is observed under lower-IS conditions. The presence of extracellular polymeric substances (EPS) was found to be the main contributor to deposition behavior. Overall, these results provide plausible evidence for the presence of amoeba in tap water. Furthermore, this is one of the first studies to examine the mechanisms affecting the fate of amoeba spores in porous media, providing a significant baseline for future research to minimize the safety risk presented by amoeba in drinking water systems.

Abstract 590: Autophagy is Required for Endothelial Atheroprotective Signaling Under Physiological Flow

Objective: Preferential development of atherosclerotic lesions in areas of low shear stress is associated with increased endothelial inflammation, apoptosis and senescence. On the contrary, high shear stress areas are protected from plaque development, but the mechanisms remain elusive. Autophagy is a protective mechanism allowing recycling of defective organelles and proteins to maintain cellular homeostasis. Our aim was to understand the role of autophagy in athero-protective effects of high shear stress. Approach and Results: We used the parallel plate chamber system in vitro to generate different shear stress conditions on endothelial cell deficient or not in autophagy (shRNA Atg5 vs. control) and examined autophagic flux in cells transfected with RFP-GFP-LC3 plasmid. Endothelial autophagy was silenced in Atg5 flox/flox ; VE-cadherin-cre mice. We first demonstrated in human and murine arteries and in cultured endothelial cells that atheroprotective shear stress activates endothelial autophagic flux. On the opposite, endothelial cells exposed to atheroprone low shear stress displayed inefficient autophagy, associated with activation of mTOR, inhibition of AMPKα pathways and blockade of the autophagic flux. Interestingly, plaque burden augmented specifically in areas usually atheroresistant in ApoE -/- hypercholesterolemic mice deficiency in endothelial autophagy. Both in cultured endothelial cells and in transgenic mice, deficiency in endothelial autophagy was associated with a defect in endothelial alignment with flow direction, a hallmark of endothelial cell health. Deficiency in endothelial autophagy increased endothelial senescence and TNFα-induced inflammation under high shear stress conditions. These effects were associated with impaired KLF2 activation. In transgenic mice, endothelial senescence and apoptosis was augmented in high shear stress areas of the descending thoracic aorta when compared to control littermate mice. Conclusions: Altogether, these results show that adequate endothelial autophagic flux under high shear stress limits atherosclerotic plaque formation by preventing inflammation, senescence and apoptosis.