<p><i>Scutellaria barbata</i> is a medicinal plant with anti-inflammatory, antioxidant, and antitumor properties. Limited studies exist on the link between <i>S. barbata</i> and liver fibrosis. The focus of this study is to examine the impact of <i>S. barbata</i>-containing serum on rat hepatocytes undergoing hepatic fibrosis. Molecular mechanisms underlying the observed effects are sought to be predicted. Transforming growth factor β1 (TGF-β1)-treated hepatic stellate cells (HSCs) supernatant was utilized to produce hepatic fibrosis-like conditions in hepatocytes BRL-3A cultured in vitro. <i>S. barbata</i>-containing serum was used as an intervention, with various dosage groups and a positive drug group (N-acetylcysteine). Cell proliferation, mitochondrial membrane potential (MMP), apoptosis, and expression of apoptosis-related proteins and genes were assessed through various assays and techniques. Bioinformatics analysis was employed to predict target genes and signaling pathways affected by <i>S. barbata</i>. Chemical components of S. barbata in the serum were detected by ultra-high-performance liquid chromatography-mass spectrometry (UHPLC-QE-MS) was used to identify. Cellular experiments demonstrated that <i>S. barbata</i>-containing serum restored cell proliferation and reduced apoptotic activity induced by the fibrosis model, with a significant downregulation of apoptosis-related proteins (cleaved-Caspase-3, Bax), a substantial upregulation of the anti-apoptotic protein BCL-2, and a substantial elevation in the level of cellular MMP. Bioinformatics analysis highlighted the involvement of <i>S. barbata</i> in hepatocyte apoptosis during liver fibrosis, possibly through pathways like PI3K-Akt. UHPLC-QE-MS identified 29 chemical components of <i>S. barbata</i> in the bloodstream, suggesting their role in anti-hepatic fibrosis effects. <i>S. barbata</i> was found to effectively inhibit hepatocyte apoptosis during hepatic fibrosis.</p>