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Neocryptotanshinone inhibits lipopolysaccharide-induced inflammation in RAW264.7 macrophages by suppression of NF- κ B and iNOS signaling pathways

View Article: PubMed Central

ABSTRACT

Neocryptotanshinone (NCTS) is a natural product isolated from traditional Chinese herb Salvia miltiorrhiza Bunge. In this study, we investigated its anti-inflammatory effects in lipopolysaccharide (LPS)-stimulated mouse macrophage (RAW264.7) cells. MTT results showed that NCTS partly reversed LPS-induced cytotoxicity. Real-time PCR results showed that NCTS suppressed LPS-induced mRNA expression of inflammatory cytokines, including tumor necrosis factor α (TNFα), interleukin-6 (IL-6) and interleukin-1β (IL-1β). Moreover, NCTS could decrease LPS-induced nitric oxide (NO) production. Western blotting results showed that NCTS could down-regulate LPS-induced expression of inducible nitric oxide synthase (iNOS), p-IκBα, p-IKKβ and p-NF-κB p65 without affecting cyclooxygenase-2 (COX-2). In addition, NCTS inhibited LPS-induced p-NF-κB p65 nuclear translocation. In conclusion, these data demonstrated that NCTS showed anti-inflammatory effect by suppression of NF-κB and iNOS signaling pathways.

No MeSH data available.


Effects of NCTS on LPS-induced production of NO2− in the cell supernatants and intracellular NO formation in RAW264.7 macrophages. Cells were treated with LPS with or without NCTS or SMT co-incubation for 12, 24 and 48 h. The NO2− contents in the cell supernatants were determined with Griess reagent (A-C); the intracellular NO formation was monitored with fluorescence probe DAF-DA (D). Data were presented as the means±SD of three independent experiments. ##P<0.01 vs. the control group; **P<0.01, *P<0.05 vs. LPS-stimulated group. NCTS, Neocryptotanshinone; Cont, Control group; SMT, S-methylisothiourea sulfate.
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f0020: Effects of NCTS on LPS-induced production of NO2− in the cell supernatants and intracellular NO formation in RAW264.7 macrophages. Cells were treated with LPS with or without NCTS or SMT co-incubation for 12, 24 and 48 h. The NO2− contents in the cell supernatants were determined with Griess reagent (A-C); the intracellular NO formation was monitored with fluorescence probe DAF-DA (D). Data were presented as the means±SD of three independent experiments. ##P<0.01 vs. the control group; **P<0.01, *P<0.05 vs. LPS-stimulated group. NCTS, Neocryptotanshinone; Cont, Control group; SMT, S-methylisothiourea sulfate.

Mentions: As shown in Fig. 4, LPS treatment induced large amount of NO2− production in cell supernatant. Furthermore, a time-dependent manner was observed (Fig. 4A-C). SMT, an iNOS inhibitor, dramatically suppressed LPS induced NO2− production after 24 h treatment. Compared with the LPS treated groups, NCTS at the concentration of 20 μmol/L remarkably reduced the NO2− production (P<0.01, 12 h; P<0.05, 24 h and 48 h) NCTS, at the concentration of 10 μmol/L decreased NO2− production at 12 h (P<0.05).


Neocryptotanshinone inhibits lipopolysaccharide-induced inflammation in RAW264.7 macrophages by suppression of NF- κ B and iNOS signaling pathways
Effects of NCTS on LPS-induced production of NO2− in the cell supernatants and intracellular NO formation in RAW264.7 macrophages. Cells were treated with LPS with or without NCTS or SMT co-incubation for 12, 24 and 48 h. The NO2− contents in the cell supernatants were determined with Griess reagent (A-C); the intracellular NO formation was monitored with fluorescence probe DAF-DA (D). Data were presented as the means±SD of three independent experiments. ##P<0.01 vs. the control group; **P<0.01, *P<0.05 vs. LPS-stimulated group. NCTS, Neocryptotanshinone; Cont, Control group; SMT, S-methylisothiourea sulfate.
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

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f0020: Effects of NCTS on LPS-induced production of NO2− in the cell supernatants and intracellular NO formation in RAW264.7 macrophages. Cells were treated with LPS with or without NCTS or SMT co-incubation for 12, 24 and 48 h. The NO2− contents in the cell supernatants were determined with Griess reagent (A-C); the intracellular NO formation was monitored with fluorescence probe DAF-DA (D). Data were presented as the means±SD of three independent experiments. ##P<0.01 vs. the control group; **P<0.01, *P<0.05 vs. LPS-stimulated group. NCTS, Neocryptotanshinone; Cont, Control group; SMT, S-methylisothiourea sulfate.
Mentions: As shown in Fig. 4, LPS treatment induced large amount of NO2− production in cell supernatant. Furthermore, a time-dependent manner was observed (Fig. 4A-C). SMT, an iNOS inhibitor, dramatically suppressed LPS induced NO2− production after 24 h treatment. Compared with the LPS treated groups, NCTS at the concentration of 20 μmol/L remarkably reduced the NO2− production (P<0.01, 12 h; P<0.05, 24 h and 48 h) NCTS, at the concentration of 10 μmol/L decreased NO2− production at 12 h (P<0.05).

View Article: PubMed Central

ABSTRACT

Neocryptotanshinone (NCTS) is a natural product isolated from traditional Chinese herb Salvia miltiorrhiza Bunge. In this study, we investigated its anti-inflammatory effects in lipopolysaccharide (LPS)-stimulated mouse macrophage (RAW264.7) cells. MTT results showed that NCTS partly reversed LPS-induced cytotoxicity. Real-time PCR results showed that NCTS suppressed LPS-induced mRNA expression of inflammatory cytokines, including tumor necrosis factor &alpha; (TNF&alpha;), interleukin-6 (IL-6) and interleukin-1&beta; (IL-1&beta;). Moreover, NCTS could decrease LPS-induced nitric oxide (NO) production. Western blotting results showed that NCTS could down-regulate LPS-induced expression of inducible nitric oxide synthase (iNOS), p-I&kappa;B&alpha;, p-IKK&beta; and p-NF-&kappa;B p65 without affecting cyclooxygenase-2 (COX-2). In addition, NCTS inhibited LPS-induced p-NF-&kappa;B p65 nuclear translocation. In conclusion, these data demonstrated that NCTS showed anti-inflammatory effect by suppression of NF-&kappa;B and iNOS signaling pathways.

No MeSH data available.