The expression of MCOLN1 in preeclampsia maintains the balance of autophagy and pyroptosis induced by reactive oxygen species.

Reactive oxygen species (ROS) can stimulate autophagy and pyroptosis, which have been slightly reported in preeclampsia (PE). Mucolipin TRP Cation Channel 1 (MCOLN1) is a sensor for autophagy to receive ROS. This study aimed to investigate the role of MCOLN1 on ROS-mediated autophagy and pyroptosis in an in vitro model of PE. HTR-8/SVneo cells were treated with different concentrations of rotenone and pre-transfected with plasmids overexpressing or silencing MCOLN1. Thiazolyl blue tetrazolium bromide was used to examine the effects of rotenone and MCOLN1 on trophoblasts. ROS levels were detected by 2',7'-Dichlorodihydrofluorescein Diacetate (DCFH-DA). The levels of MCOLN1, pyroptosis and autophagy-related proteins were determined by western blot, and the mRNA level of MCOLN1 was examined by quantitative real-time polymerase chain reaction. The effect of MCOLN1 on rotenone-treated trophoblast cells was also analyzed using flow cytometry, wound healing, Transwell, and Enzyme-linked immunosorbent assay (ELISA) experiments. Rotenone decreased cell viability and induced ROS levels in a concentration-dependent manner in HTR-8/SVneo. MCOLN1 overexpression partially reversed the inhibition of cell viability, migration and invasion and the promotion of apoptosis by rotenone; but MCOLN1 silencing enhanced the effect of rotenone. Unlike MCOLN1 silencing, MCOLN1 overexpression reduced rotenone-elevated Interleukin (IL)-18, IL-1β, Gasdermin D (GSDMD), nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), pro-caspase 1, caspase 1, but enhanced Beclin 1, Microtubule-associated protein 1 light chain 3 (LC3)II/LC3I levels, and inhibited Sequestosome 1 (P62) expression suppressed by rotenone. MCOLN1 mediates autophagy and pyroptosis balance to alleviate trophoblast dysfunction caused by excess ROS.