New sight into interaction between endoplasmic reticulum stress and autophagy induced by vanadium in duck renal tubule epithelial cells.

Vanadium (V) is a common environmental and industrial pollutant that can cause nephrotoxicity in animals in excess. The purpose of this research was to explore the interaction between endoplasmic reticulum (ER) stress and autophagy induced by V in the kidney of ducks. Duck renal tubule epithelial cells were exposed to different concentrations of sodium metavanadate (NaVO3) (0, 100 and 200 µM) and PERK inhibitor (GSK, 1 µM), or autophagy inhibitor (chloroquine, 50 µM) alone for 24 h (chloroquine for the last 4 h). The results showed that exposure to V caused the dilatation and swelling of the ER and intracellular calcium overload, and upregulated PERK, eIF2α, ATF4 and CHOP mRNA levels and p-PERK and CHOP protein levels associated with ER stress in cells. Additionally, V markedly increased the number of autophagosomes, acidic vesicular organelles (AVOs) and LC3 puncta, as well as the mRNA levels of Beclin1, Atg5, Atg12, LC3A and LC3B and protein levels of Beclin1, Atg5 and LC3B-II/LC3B-I, but decreased the imRNA and protein levels of p62. Moreover, treatment with the PERK inhibitor ameliorated the changed factors above induced by V, but the V-induced variation of ER-stress related factors were aggravated after treatment with the autophagy inhibitor. Together, our data suggested that excessive V could induce ER stress and autophagy in duck renal tubular epithelial cells. ER stress might promote V-induced autophagy via the PERK/ATF4/CHOP signaling pathway, and autophagy may play a role in alleviating ER stress induced by V.

Endoplasmic reticulum-mitochondria coupling attenuates vanadium-induced apoptosis via IP3R in duck renal tubular epithelial cells.

Vanadium (V) is necessary for the health and growth of animals, but excessive V has harmful effects on the ecosystem health. Endoplasmic reticulum (ER)-mitochondria coupling as a membrane structure connects the mitochondrial outer membrane with the ER. The mitochondria-associated ER membrane (MAM) is a region of the ER-mitochondria coupling and is essential for normal cell function. Currently, the crosstalk between ER-mitochondrial coupling and apoptosis in the toxic mechanism of V on duck kidney is still unclear. In this study, duck renal tubular epithelial cells were incubated with different concentrations of sodium metavanadate (NaVO3) and/or inositol triphosphate receptor (IP3R) inhibitor 2-aminoethyl diphenyl borate (2-APB) for 24 h. The results showed that V could significantly increase lactate dehydrogenase (LDH) release, the mitochondrial calcium level and the numbers of the fluorescent signal points of IP3R; shortened the length ER-mitochondria coupling and reduced its formation; markedly upregulate the mRNA levels of MAM-related genes and protein levels, causing MAM dysfunction. Additionally, V treatment appeared to upregulate pro-apoptotic genes and downregulate anti-apoptotic genes, followed by cell apoptosis. The V-induced changes were alleviated by treatment with IP3R inhibitor. In summary, V could induce the dysfunction of ER-mitochondrial coupling and apoptosis, and inhibition of ER-mitochondrial coupling could attenuate V-induced apoptosis in duck renal tubular epithelial cells.