Journal
PLOS ONE
Volume 9, Issue 7, Pages -Publisher
PUBLIC LIBRARY SCIENCE
DOI: 10.1371/journal.pone.0103229
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Funding
- National Science Council [NSC99-2628-B-077-001-MY3, NSC-102-2320-B-077-001]
- National Research Institute of Chinese Medicine in Taiwan [NRICM99-DBCM-05]
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Background: Anti-inflammation via inhibition of NF-kappa B pathways in hepatic stellate cells (HSCs) is one therapeutic approach to hepatic fibrosis. Tanshinone IIA (C19H18O3, Tan IIA) is a lipophilic diterpene isolated from Salvia miltiorrhiza Bunge, with reported anti-inflammatory activity. We tested whether Tan IIA could inhibit HSC activation. Materials and Methods: The cell line of rat hepatic stellate cells (HSC-T6) was stimulated with lipopolysaccharide (LPS) (100 ng/ml). Cytotoxicity was assessed by MTT assay. HSC-T6 cells were pretreated with Tan IIA (1, 3 and 10 mu M), then induced by LPS (100 ng/ml). NF-kappa B activity was evaluated by the luciferase reporter gene assay. Western blotting analysis was performed to measure NF-kappa B-p65, and phosphorylations of MAPKs (ERK, JNK, p38). Cell chemotaxis was assessed by both wound-healing assay and trans-well invasion assay. Quantitative real-time PCR was used to detect gene expression in HSC-T6 cells. Results: All concentrations of drugs showed no cytotoxicity against HSC-T6 cells. LPS stimulated NF-kappa B luciferase activities, nuclear translocation of NF-kappa B-p65, and phosphorylations of ERK, JNK and p38, all of which were suppressed by Tan IIA. In addition, Tan IIA significantly inhibited LPS-induced HSCs chemotaxis, in both wound-healing and trans-well invasion assays. Moreover, Tan IIA attenuated LPS-induced mRNA expressions of CCL2, CCL3, CCL5, IL-1 beta, TNF-alpha, IL-6, ICAM-1, iNOS, and alpha-SMA in HSC-T6 cells. Conclusion: Our results demonstrated that Tan IIA decreased LPS-induced HSC activation.
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