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Inhibitory effect of tanshinone IIA on rat hepatic stellate cells.

Liu YW, Huang YT - PLoS ONE (2014)

Bottom Line: LPS stimulated NF-κB luciferase activities, nuclear translocation of NF-κB-p65, and phosphorylations of ERK, JNK and p38, all of which were suppressed by Tan IIA.Moreover, Tan IIA attenuated LPS-induced mRNA expressions of CCL2, CCL3, CCL5, IL-1β, TNF-α, IL-6, ICAM-1, iNOS, and α-SMA in HSC-T6 cells.Our results demonstrated that Tan IIA decreased LPS-induced HSC activation.

View Article: PubMed Central - PubMed

Affiliation: Institute of Traditional Medicine, School of Medicine, National Yang-Ming University, Taipei, Taiwan.

ABSTRACT

Background: Anti-inflammation via inhibition of NF-κB 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 µM), then induced by LPS (100 ng/ml). NF-κB activity was evaluated by the luciferase reporter gene assay. Western blotting analysis was performed to measure NF-κB-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-κB luciferase activities, nuclear translocation of NF-κB-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-1β, TNF-α, IL-6, ICAM-1, iNOS, and α-SMA in HSC-T6 cells.

Conclusion: Our results demonstrated that Tan IIA decreased LPS-induced HSC activation.

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Related in: MedlinePlus

Chemical structures and cytotoxicity of salvianolic acid B and tanshinone IIA.Chemical structures of Sal B (A) and Tan IIA (B). (C) MTT assay was performed to assess HSC-T6 cells viability with Tan IIA (1, 3, and 10 µM) and Sal B (200 µM) treatment for 24 hr. There was no cytotoxicity in all concentrations of drugs. n = 3.
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pone-0103229-g001: Chemical structures and cytotoxicity of salvianolic acid B and tanshinone IIA.Chemical structures of Sal B (A) and Tan IIA (B). (C) MTT assay was performed to assess HSC-T6 cells viability with Tan IIA (1, 3, and 10 µM) and Sal B (200 µM) treatment for 24 hr. There was no cytotoxicity in all concentrations of drugs. n = 3.

Mentions: Both Sal B and Tan IIA (Figure 1A and B) are key constituents of S. miltiorrhiza, the former coming from aqueous extract and the latter being in lipophilic part. According to our previous studies, Sal B has already been found to protect hepatic function, such as inhibiting CCl4 induced ROS accumulation in hepatocytes and HSC activation [21], [23]. Therefore, we further studied bioactivities of Tan IIA against HSC activation. Using MTT assays, neither Tan IIA (1, 3 and 10 µM) nor Sal B (200 µM) showed cytotoxicity to HSC-T6 cells for 24 hr (Figure 1C). To confirm whether there was proliferative effect of Tan IIA and Sal B in HSC-T6 cells, we used immunofluorescent staining of Ki67. The results show that there was no increase in proliferation by Tan IIA or Sal B (Figure S1).


Inhibitory effect of tanshinone IIA on rat hepatic stellate cells.

Liu YW, Huang YT - PLoS ONE (2014)

Chemical structures and cytotoxicity of salvianolic acid B and tanshinone IIA.Chemical structures of Sal B (A) and Tan IIA (B). (C) MTT assay was performed to assess HSC-T6 cells viability with Tan IIA (1, 3, and 10 µM) and Sal B (200 µM) treatment for 24 hr. There was no cytotoxicity in all concentrations of drugs. n = 3.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4116159&req=5

pone-0103229-g001: Chemical structures and cytotoxicity of salvianolic acid B and tanshinone IIA.Chemical structures of Sal B (A) and Tan IIA (B). (C) MTT assay was performed to assess HSC-T6 cells viability with Tan IIA (1, 3, and 10 µM) and Sal B (200 µM) treatment for 24 hr. There was no cytotoxicity in all concentrations of drugs. n = 3.
Mentions: Both Sal B and Tan IIA (Figure 1A and B) are key constituents of S. miltiorrhiza, the former coming from aqueous extract and the latter being in lipophilic part. According to our previous studies, Sal B has already been found to protect hepatic function, such as inhibiting CCl4 induced ROS accumulation in hepatocytes and HSC activation [21], [23]. Therefore, we further studied bioactivities of Tan IIA against HSC activation. Using MTT assays, neither Tan IIA (1, 3 and 10 µM) nor Sal B (200 µM) showed cytotoxicity to HSC-T6 cells for 24 hr (Figure 1C). To confirm whether there was proliferative effect of Tan IIA and Sal B in HSC-T6 cells, we used immunofluorescent staining of Ki67. The results show that there was no increase in proliferation by Tan IIA or Sal B (Figure S1).

Bottom Line: LPS stimulated NF-κB luciferase activities, nuclear translocation of NF-κB-p65, and phosphorylations of ERK, JNK and p38, all of which were suppressed by Tan IIA.Moreover, Tan IIA attenuated LPS-induced mRNA expressions of CCL2, CCL3, CCL5, IL-1β, TNF-α, IL-6, ICAM-1, iNOS, and α-SMA in HSC-T6 cells.Our results demonstrated that Tan IIA decreased LPS-induced HSC activation.

View Article: PubMed Central - PubMed

Affiliation: Institute of Traditional Medicine, School of Medicine, National Yang-Ming University, Taipei, Taiwan.

ABSTRACT

Background: Anti-inflammation via inhibition of NF-κB 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 µM), then induced by LPS (100 ng/ml). NF-κB activity was evaluated by the luciferase reporter gene assay. Western blotting analysis was performed to measure NF-κB-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-κB luciferase activities, nuclear translocation of NF-κB-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-1β, TNF-α, IL-6, ICAM-1, iNOS, and α-SMA in HSC-T6 cells.

Conclusion: Our results demonstrated that Tan IIA decreased LPS-induced HSC activation.

Show MeSH
Related in: MedlinePlus