MiR-130b-3p regulates steroidogenesis-related markers, cell viability, inflammation, oxidative stress, and pyroptosis-associated responses by targeting PPARG in LPS-exposed goat granulosa cells.

Abstract

LPS, an endotoxin derived from Gram-negative bacteria, disrupts granulosa cell (GCs) function, and impairs ovarian activity. This study aimed to investigate the role of miR-130b-3p targeting PPARG expression in LPS-exposed goat GCs. Results indicated that LPS decreased PPARG expression, while increased the level of miR-130b-3p which binds to the 3'UTR of PPARG mRNA. PPARG knockdown and miR-130b-3p overexpression further aggravated the reductions in PPARG expression and cell viability under LPS stimulation, while PPARG overexpression and miR-130b-3p inhibition effectively alleviated these LPS-induced inhibitory effects. LPS decreased the mRNA abundances of HSD3B, CYP19A1, CDK4, PCNA, SOD, and CAT, as well as the protein levels of HSD3B and CDK4; Conversely, it increased the mRNA abundances of IL-1β, TNF-α, Caspase-1, and GSDMD, as well as IL-1β protein and intracellular ROS levels. These LPS-induced alterations were further intensified by si-PPARG or miR-130b-3p mimics, as evidenced by more pronounced changes in HSD3B, CYP19A1, CDK4, SOD, CAT, IL-1β, TNF-α, and Caspase-1 and GSDMD mRNA abundances, as well as HSD3B (except siRNA) and CDK4 protein, and ROS levels. In contrast, PPARG overexpression or miR-130b-3p inhibition alleviated LPS-induced changes in HSD3B, CYP19A1, CDK4, PCNA, SOD, and CAT, IL-1β, TNF-α, Caspase-1 and GSDMD at the mRNA abundances, as well as HSD3B and CDK4 protein, and ROS levels. In conclusion, miR-130b-3p modulates steroidogenesis-related markers, cell viability, inflammation, oxidative stress, and pyroptosis-associated responses by modulating PPARG expression in goat GCs exposed to LPS.