Chrysanthemum indicum ethanol extract attenuates hepatic stellate cell activation in vitro and thioacetamide-induced hepatofibrosis in rats

Abstract

Objective: To investigate the antifibrotic effects of Chrysanthemum indicum ethanol extract (CIEE) against activated hepatic stellate cells (HSC) and thioacetamide (TAA)-induced hepatofibrosis in rats. Methods: Cell viability and proliferation of HSC-T6 cells were measured using MTT assay. Primary HSCs were used to study morphology. TAA (200 mg/kg) was used to induced hepatic fibrosis in rats. CIEE (100 and 500 mg/kg) and silymarin (50 mg/kg) were administered orally. Liver functions including alanine transaminase, aspartate transaminase, glutathione, and hydroxyproline levels were measured using commercial kits. Liver sections and fibrotic biomarker expression were measured using hematoxylin and eosin staining and real-time polymerase chain reaction. Results: In vitro study revealed that CIEE (0.1, 0.25, and 0.5 mg/mL) inhibited the proliferation of activated HSCs exposed to transforming growth factor (TGF)-β and restored the activated primary HSC morphology. In in vivo studies, TAA-induced increase in liver/body weight ratio (5.46 ± 0.26) was significantly reduced (4.13 ± 0.22) by CIEE (P<0.05 at 500 mg/kg). CIEE (100 and 500 mg/kg) improved the liver functions by significantly attenuating changes in alanine transaminase, aspartate transaminase, glutathione, and hydroxyproline levels (P<0.05). Further, CIEE (100 and 500 mg/kg) ameliorated the histological changes in liver tissue and TGF-β expression significantly (P<0.05) in TAA-induced rats. Conclusions: CIEE significantly protects against TAA-induced liver damage in rats and can be used in the treatment of liver fibrosis.

Keywords

• chrysanthemum indicum; fibrosis; hepatoprotective; hydroxyproline; thioacetamide