Metabolic dysfunction-associated steatotic liver disease (MASLD) encompasses a spectrum of progressive liver disorders that, if unresolved, can trigger inflammation and fibrosis. Hepatic stellate cells (HSCs) are key mediators of these fibrotic processes in the liver, undergoing activation in response to liver injury or chronic inflammation. Thereby, quiescent HSCs transdifferentiate into proliferative and migratory myofibroblast-like cells that secrete large quantities of extracellular matrix (ECM) proteins. This phenotypic shift is highly energy demanding, provoking metabolic reprogramming of activated HSCs. The increase in metabolic activity elevates oxidative stress and disrupts cellular redox homeostasis. Recent advances in therapy of MASLD related conditions focus on inhibiting HSC activation, including approaches targeting stearoyl-CoA desaturase (SCD), the rate-limiting enzyme responsible for the biosynthesis of monounsaturated fatty acids. Pharmacological inhibition and genetic knockdown of SCD have been associated with reduced activation of HSCs in vitro, yet the underlying mechanisms that link diminished SCD activity to these antifibrotic effects in HSCs remain only partially defined. To address this, we setup an in vitro model to characterize HSC activation at the protein level. Using the widely employed immortalized human HSC line LX-2, we compared the proteomic profiles of pseudo-quiescent and TGF-β-activated cells. To delineate the mechanisms underlying reduced HSC activation upon loss of SCD activity, we specifically inhibited SCD with the small-molecule inhibitor A939572 and assessed its effects in both pseudo-quiescent and activated states. We validated our in vitro model of HSC activation by demonstrating robust upregulation of fibrogenic marker proteins and increased ECM secretion upon TGF-β stimulation. Activated LX-2 cells also showed elevated SCD expression underlining its importance in the activation process. Inhibition of SCD markedly reduced key fibrogenic markers, including α-SMA, COL1A1, MMP2, TGFB2 and FN1, demonstrating a strong suppression of HSC activation. Impaired SCD activity also caused pronounced alterations in redox-regulatory pathways, particularly within the transsulfuration and glutathione pathway, indicating its direct impact on cellular oxidative stress defence within HSCs. These findings provide a step towards elucidating the protective and anti-fibrotic effect of SCD inhibition in HSCs and corroborate therapeutic potential of SCD inhibition for targeted fibrosis therapy.