Nutr Res Pract.  2017 Dec;11(6):470-478. 10.4162/nrp.2017.11.6.470.

Anti-fibrotic effects of Orostachys japonicus A. Berger (Crassulaceae) on hepatic stellate cells and thioacetamide-induced fibrosis in rats

Affiliations
  • 1Department of Biotechnology, College of Biomedical and Health Sciences, Konkuk University, 268 Chungwon-daero, Chungju-si, Chungbuk 27478, Korea. minds@kku.ac.kr
  • 2Department of Applied Life Science, Graduate School of Konkuk University, 268 Chungwon-daero, Chungju-si, Chungbuk 27478, Korea.
  • 3Food One Corp., 260 Sincheoksandan 5-ro, Deoksan-myeon, Jincheon-gun, Chungbuk 27850, Korea.
  • 4Department of Internal Medicine, School of Medicine, Konkuk University, Chungju, Chungbuk 27376, Korea.
  • 5R&D center, Korean Drug Co., Ltd., Seoul 06300, Korea.

Abstract

BACKGROUND
/OBJECTIVE: Orostachys 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/METHODS
An 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.
RESULTS
OJE (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.
CONCLUSIONS
OJE can be developed as a potential agent for the treatment of hepatofibrosis.

Keyword

Liver; glutathione; hydroxyproline; apoptosis; silymarin
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