| Summary: | 碩士 === 國立陽明大學 === 傳統醫藥學研究所 === 97 === Anti-inflammation via inhibition of NF-kB pathways in hepatic stellate cells (HSCs) is an important therapeutic approach to hepatic fibrosis. Tanshinone IIA and cryptotanshinone are two major diterpenes from Salvia miltiorrhiza Bunge, with reported anti-inflammatory activity. We tested whether tanshinone IIA or cryptotanshinone could inhibit HSC activation via NF-kB.
A cell line of rat hepatic stellate cells (HSC-T6) was stimulated with tumor necrosis factor (TNF)-a (1 ng/ml) or lipopolysaccharide (LPS) (100 ng/ml). Cytotoxicity was assessed by MTT assay. NF-kB activity was assessed by the luciferase reporter gene assay. Western blot analysis was performed to measure NF-kB-p65 and AP-1-JunD nuclear translocation, and phosphorylations of MAPKs (ERK, JNK, and p38). The mRNA expression of iNOS gene was detected by real-time PCR. HSC-T6 cells were pretreated with tanshinone IIA and cryptotanshinone, then induced by TNF-a and LPS. Both cryptotanshinone (10 uM) and tanshinone IIA (3 and 10 uM) inhibited NF-kB luciferase activity. Tanshinone IIA (10 uM) also inhibited LPS- and TNF-a-stimulated both NF-kB-p65 nuclear translocation and AP-1-JunD nuclear translocation. However, tanshinone IIA did not affect the phosphorylations of ERK, JNK and p38. Tanshinone IIA inhibit TNF-a-stimulated mRNA expression of iNOS, but not LPS-stimulated. Tanshinone IIA was not cytotoxic at 3 and 10 M. In conclusion, our results suggested that tanshinone IIA inhibited LPS- and TNF-a-stimulated NF-kB activation in HSCs.
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