The current study examines the role of testosterone and androgen deprivation in regulating mitochondrial quality control, autophagy, and apoptosis during CCl4-induced chronic liver injury. Male rats were allocated to Sham, CCl4, TP + CCl4 (testosterone-treated), and Cas + CCl4 (castrated) groups for 4, 8, and 12 weeks, validated through in vitro assays. Mitochondrial function (MitoTracker, MFN2), lysosomal integrity (LAMP1), autophagy markers (LC3B, Beclin-1, p62/SQSTM1), apoptosis regulators (Bcl-2, cleaved caspase-3), and mTOR signaling were assessed by qRT-PCR, immunohistochemistry, and immunofluorescence. CCl4 exposure caused progressive increases in cellular stress and fibrotic markers (α-SMA, TGF-β1, and TXNIP), progressive mitochondrial dysfunction, impaired mitochondrial-lysosomal coordination, reduced Beclin-1 and LC3B, p62 accumulation, and a shift toward an anti-apoptotic Bcl-2 imbalance. Testosterone treatment restored mitochondrial membrane potential in vitro, normalized MitoTracker and MFN2 expression, preserved LAMP1 levels, and partially sustained mitochondrial-lysosomal coupling. TP also enhanced Beclin-1 and LC3B, reduced p62 accumulation, and sustained mTOR activity, consistent with improved autophagic flux. Moreover, TP-lowered Bcl-2 was associated with controlled cleaved caspase-3 activation, suggesting selective clearance of damaged hepatocytes rather than widespread apoptosis. In contrast, Cas + CCl4 animals showed exaggerated mitochondrial impairment, persistent p62 accumulation, reduced LC3B and mTOR signaling, and a pro-apoptotic cleaved caspase-3 profile. Together, these findings demonstrate that testosterone supports mitochondrial-lysosomal homeostasis and autophagy during toxic liver injury, whereas androgen deprivation favors defective autophagy and maladaptive apoptosis, underscoring hormone-dependent regulation of hepatocellular stress responses.
