Anti-fibrotic effects ofOrostachys japonicusA. Berger (Crassulaceae) on hepatic stellate cells and thioacetamide-induced fibrosis in rats

Abstract

BACKGROUND/OBJECTIVEOrostachys japonicus A. Berger (Crassulaceae) has been used in traditional herbal medicines in Korea and other Asian countries to treat various diseases, including liver disorders. In the present study, the anti-fibrotic effects of O. japonicus extract (OJE) in cellular and experimental hepatofibrotic rat models were investigated.MATERIALS/METHODSAn in vitro hepatic stellate cells (HSCs) system was used to estimate cell viability, cell cycle and apoptosis by MTT assay, flow cytometry, and Annexin V-FITC/PI staining techniques, respectively. In addition, thioacetamide (TAA)-induced liver fibrosis was established in Sprague Dawley rats. Briefly, animals were divided into five groups (n = 8): Control, TAA, OJE 10 (TAA with OJE 10 mg/kg), OJE 100 (TAA with OJE 100 mg/kg) and silymarin (TAA with Silymarin 50 mg/kg). Fibrosis was induced by treatment with TAA (200 mg/kg, i.p.) twice per week for 13 weeks, while OJE and silymarin were administered orally two times per week from week 7 to 13. The fibrotic related gene expression serum biomarkers glutathione and hydroxyproline were estimated by RT-PCR and spectrophotometry, respectively, using commercial kits.RESULTSOJE (0.5 and 0.1 mg/mL) and silymarin (0.05 mg/mL) treatment significantly (P < 0.01 and P < 0.001) induced apoptosis (16.95% and 27.48% for OJE and 25.87% for silymarin, respectively) in HSC-T6 cells when compared with the control group (9.09%). Further, rat primary HSCs showed changes in morphology in response to OJE 0.1 mg/mL treatment. In in vivo studies, OJE (10 and 100 mg/kg) treatment significantly ameliorated TAA-induced alterations in levels of serum biomarkers, fibrotic related gene expression, glutathione, and hydroxyproline (P < 0.05-P < 0.001) and rescued the histopathological changes.CONCLUSIONSOJE can be developed as a potential agent for the treatment of hepatofibrosis.

Aster tataricus Linn., Suppresses Hepatic Stellate Cell Activation and Protects Against Thioacetamide-induced Liver Fibrosis in Rats

“…HSC-T6 cells were cultured in Dulbecco`s modified Eagle media (DMEM, Invitrogen, Carlsbad, CA, USA) supplemented with 5 % fetal bovine serum (FBS, Invitrogen, Carlsbad, CA, USA) and 1 % antibiotic-antimycotic (Invitrogen, Carlsbad, CA, USA) in a humidified atmosphere of 5% CO2 at 37°. The Chang liver cell line was used as a normal human cell line derived from normal liver tissue and cultured as described previously [26] . HSC-T6 cells were activated by serum starvation before treatment with ATE.…”

“…The effect of ATE on overall cell viability was evaluated using MTT assay and the results were measured on an ELISA reader at 540 nM as described previously [26] . Briefly, the cells were treated with ATE (24 h) at various concentrations (0, 0.1, 0.5, 1.0 mg/ml) and were incubated with 0.5 mg/ml MTT for 3 h, and then the reaction was interrupted by addition of DMSO.…”

“…Cell cycle analysis in HSC-T6 cells was evaluated using ATE at 0.1 and 0.5 mg/ml concentrations measured by flow cytometer (FACSCalibur, BD Biosciences, San Jose, CA, USA) using a method reported previously [26] . For apoptosis, Annexin V-FITC and PI apoptosis detection kit I was used according to manufacturer's instructions.…”

“…The protocol for qRT-PCR was followed as described previously [26] . Briefly, total RNA was extracted from tissue samples using TRIzol Plus RNA Purification Kit (Qiagen, Valencia, CA, USA).…”

Studies on the Anti‐Angiogenic Activities of Wild and Cultivated Orostachys japonicus Extracts in Human Umbilical Vein Endothelial Cells

“…O. japonicus , which has traditionally been used as a food ingredient and herbal remedy in Asia to treat various diseases, lives in niche environments such as on mountain rocks or roof tiles. Numerous studies have reported that O. japonicus extracts have biological effects on cancer (Kim et al., ; Won et al., ), inflammation (Lee, Ryu, Lee, & Lee, ), and fibrosis (Koppula et al., ). In addition, they contain various phenolic compounds, including gallic acid (Hur & Park, ), triterpenoids (Je Ma, Jung, Lee, Kim, & Sung, ), and flavonoids (Lee et al., ).…”