Obtusifolin mitigates CCl4-induced inflammation and liver fibrosis by modulating the ROCK2/ Cofilin-1 / NFκB pathway

Liver fibrosis is a progressive clinical condition characterised by an abnormal accumulation of extracellular matrix (ECM) components, which are predominantly generated by activated hepatic stellate cells (HSCs) and Other fibrogenic cells. This process leads to liver stiffness, which can progress to cirrhosis, an irreversible condition, and ultimately result in liver failure. Obtusifolin (OBT), an anthraquinone derivative isolated from the seeds of Senna obtusifolia, is known for its potent anti-inflammatory and antioxidant properties. The present study evaluated the anti-fibrotic efficacy of OBT using in vitro models, including TGFβ-induced differentiation in HUCLS, HSC-LX2, and HHSECs, and an in vivo CCl₄-induced liver fibrosis model in mice. The in vitro results demonstrated that OBT significantly suppressed the TGFβ-induced upregulation of fibrotic markers at both mRNA and protein levels. In vivo, findings showed that CCl₄ treatment caused significant increases in serum liver function markers, inflammatory cell infiltration, Collagen deposition, and septal fibrosis in liver tissues. In contrast, OBT treatment attenuated these pathological changes in a dose-dependent manner. Furthermore, assays on oxidative stress indicators revealed that OBT significantly reduced oxidative stress markers and enhanced antioxidant levels. Gene and protein expression analyses showed that OBT suppressed the CCl₄-induced elevation of inflammatory cytokines, chemokines, and fibrotic markers compared to the CCl₄ control group. Mechanistic studies demonstrated that CCl₄ administration activated the ROCK2/p-cofilin-1/pNFκB signaling pathway, whereas OBT treatment significantly modulated these molecular pathways. OBT alleviates liver inflammation and fibrosis by inhibiting oxidative stress, Collagen accumulation, and the ROCK2/NFκB pathway, highlighting its potential as a therapeutic candidate for liver fibrosis.

Cofilin-1; Inflammation; Liver injury and NFκB; Myofibroblasts; ROCK-2.