Abstract | AIM OF THE STUDY:
Icaritin is an active ingredient extracted from the plant Herba Epimedium Sagittatum (Sieb. et Zucc.) Maxim. The purpose of this study is to investigate the effects and mechanisms of icaritin-induced cell death in activated hepatic stellate cells (HSCs) and ameliorating the development of liver fibrosis in rats. MATERIALS AND METHODS: : In vitro, icaritin-induced cell death rates in HSC-T6 (rat) and LX-2 (human) HSC lines as well as normal hepatocyte cell lines HL-7702 (L02) and WRL-68 were assayed by MTT method, and the apoptotic ratios were detected by both flow cytometry and the Annexin-V-FITC Apoptosis Detection Kit. A Whole Rat Genome Microarray Kit was used to identify expression of interest genes through fold-change screening. In vivo study, experimental liver fibrosis models were built by carbon tetrachloride (CCl(4)) or common bile duct ligation (CBDL) in Wistar rats. Icaritin (1mg/kg/day, three days a week) was administered by gastric gavage for four weeks (n=6 per group). At the end of the study, serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT) as well as the contents of hydroxyproline and collagen I in liver tissues were measured. Histopathological changes and the distribution of activated HSCs were observed in the liver tissues using hematoxyline- eosin (HE) staining and immunohistochemical staining for α-smooth muscle actin (α-SMA). RESULTS:
Icaritin induced apoptosis in HSC-T6 and LX-2 in a concentration- and time-dependent manner with little toxicity to normal hepatocyte cell lines. The IC(50) of icaritin in HSC-T6 was 12.83 μM at 48 h. Apoptotic ratio of HSC-T6 treated with 24 μM icaritin was 20.19%, and the G2 phase of the cell cycle did not occur (P<0.05). Gene analysis showed that icaritin up-regulated Bak-1, Bmf and Bax expression while significantly down-regulated Bcl-2 expression (vs. control group, P<0.01). These results suggested that mitochondrial pathway played an important role in icaritin-induced apoptosis in activated HSCs. In vivo results showed that icaritin reduced the number of activated HSCs, and brought the elevated levels of AST, ALT, hydroxyproline and collagen I to normal or near normal values (vs. model group, P<0.05). CONCLUSIONS:
Icaritin can induce cell death in activated HSCs through mitochondria-mediated apoptosis, ameliorate the progression of hepatic fibrosis in rats, and could be a promising drug for treating liver fibrosis.
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Authors | Jing Li, Peng Liu, Ruixiu Zhang, Lu Cao, Haihua Qian, Jian Liao, Wen Xu, Mengchao Wu, Zhengfeng Yin |
Journal | Journal of ethnopharmacology
(J Ethnopharmacol)
Vol. 137
Issue 1
Pg. 714-23
(Sep 01 2011)
ISSN: 1872-7573 [Electronic] Ireland |
PMID | 21726622
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Copyright | Copyright © 2011 Elsevier Ireland Ltd. All rights reserved. |
Chemical References |
- Apoptosis Regulatory Proteins
- Collagen Type I
- Flavonoids
- Aspartate Aminotransferases
- Alanine Transaminase
- Hydroxyproline
- icaritin
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Topics |
- Alanine Transaminase
(blood)
- Animals
- Apoptosis
(drug effects, genetics)
- Apoptosis Regulatory Proteins
(metabolism)
- Aspartate Aminotransferases
(blood)
- Cell Line
- Cell Proliferation
(drug effects)
- Collagen Type I
(metabolism)
- Dose-Response Relationship, Drug
- Flavonoids
(pharmacology, toxicity)
- Gene Expression Profiling
- Gene Expression Regulation
(drug effects)
- Hepatic Stellate Cells
(drug effects, metabolism, pathology)
- Humans
- Hydroxyproline
(metabolism)
- Immunohistochemistry
- Inhibitory Concentration 50
- Liver
(drug effects, metabolism, pathology)
- Liver Cirrhosis, Experimental
(genetics, metabolism, pathology, prevention & control)
- Mitochondria, Liver
(drug effects, metabolism, pathology)
- Rats
- Rats, Wistar
- Time Factors
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