Search

Autophagy is a dynamic process for waste disposal and cell equilibrium. Previous studies have demonstrated that atmospheric particulate matter (APM) induces autophagy and enhances LC3II expression in human vascular endothelial cells. However, the underlying mechanism of autophagosome accumulation in human vascular endothelial cells under the exposure to APM has not been understood. In principle, the upregulation of LC3II or autophagosomes accumulation is presumably caused by the enhancement of autophagic ability, or alternatively, by the abnormal autophagic degradation. Therefore, in the current study, autophagic ability and autophagic flux are systemically studied to decipher the exact cause of autophagosomes accumulation in human umbilical vein endothelial cells (HUVECs) in response to a standard urban particulate matter, PM SRM1648a. As a result, it was observed that after 24 h of exposure, PM SRM1648a significantly increases LC3II expression with apparent autophagosomes accumulation in HUVECs. Compared with the control group, there is a time-dependent increase in p62, a protein of autophagic substrate that can be preliminarily used to evaluate the autophagic degradation, in the PM SRM1648a-exposed HUVECs, which suggested that normal function of autophagic degradation was probably impaired. Additionally, mRFP-GFP-LC3 assay and LAMP-2/LC3B co-localization suggested that autolysosomes (fusion between autophagosomes and lysosomes) were partially inhibited in PM SRM1648a-treated HUVECs. Furthermore, LC3II turn-over assay hinted that after 24 h, LC3II upregulation is attributed to the blockage of autophagic flux instead of the enhancement of autophagic induction. Mechanistically, the blockade of autophagic flux can be explained by the detrimental effects of PM SRM1648a on lysosomal function, including lysosomal destabilization, lysosomal alkalization and hydrolase inactivation, which are involved in the blockade of fusion between autophagosomes and lysosomes, further disrupting autophagic degradation and waste disposal. These observations provide evidence that PM SRM1648a destroys the equilibrium of lysosomal stability and thus results in the dysfunction of autophagic flux, eventually contributing to endothelial cell damage. Unlabelled Image • After 24 h, autophagic flux was blocked in HUVECs following exposure to PM SRM1648a. • PM SRM1648a causes defective autophagy in HUVECs. • Lysosomes are damaged in HUVECs in response to PM SRM1648a. • Particles-induced lysosomal impairment is responsible for dysfunction of autophagic flux. [ABSTRACT FROM AUTHOR]