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. 2018 Mar 21;23(4):717.
doi: 10.3390/molecules23040717.

Heterophyllin B Ameliorates Lipopolysaccharide-Induced Inflammation and Oxidative Stress in RAW 264.7 Macrophages by Suppressing the PI3K/Akt Pathways

Chunjing Yang  1 , Longtai You  2 , Xingbin Yin  3 , Yi Liu  4 , Xin Leng  5 , Wenping Wang  6 , Na Sai  7 , Jian Ni  8
Affiliations

Heterophyllin B Ameliorates Lipopolysaccharide-Induced Inflammation and Oxidative Stress in RAW 264.7 Macrophages by Suppressing the PI3K/Akt Pathways

Chunjing Yang et al. Molecules. .

Abstract

Heterophyllin B (HB), an active cyclic peptide, is a compound existing in the ethyl acetate extract of Pseudostellaria heterophylla (Miq.) Pax and exhibited the activity of inhibiting the production of NO and cytokines, such as IL-1β and IL-6, in LPS-stimulated RAW 264.7 macrophages. In addition, HB suppressed the production of ROS and the apoptosis induced by LPS in RAW 264.7 macrophages. The underlying mechanism was investigated in the LPS-induced RAW 264.7 cells. The results showed that HB decreased the level of IL-1β and IL-6 expression by qRT-PCR analysis. HB up-regulated the relative ratio of p-AKT/AKT and p-PI3K/PI3K as indicated by western blot analysis. In summary, HB inhibited the LPS-induced inflammation and apoptosis through the PI3K/Akt signaling pathways and represented a potential therapeutic target for treatment of inflammatory diseases.

Keywords: Heterophyllin B; anti-apoptosis; anti-inflammatory effect; anti-oxidative effect.

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Conflict of interest statement

The authors declare that there are no conflict of interest.

Figures

Figure 1
Figure 1
Chemical structure of Heterophyllin B (HB).
Figure 2
Figure 2
Effects of HB on the cell viability in RAW 264.7 Cells. RAW 264.7 Cells were cultured with different concentrations of HB (0, 25, 50, and 100 μM) for 24 h. The cell viability was detected by MTT assay. The values are presented as means ± SD of three independent experiments.
Figure 3
Figure 3
Effects of HB on LPS-induced NO production in RAW 264.7 Cells. RAW 264.7 Cells were pretreated with different concentrations of HB (25, 50, and 100 μM) for 1 h before treatment with 1 μg/mL LPS. After incubation for 18 h, the NO production analysis was detected by Griess test. The values are presented as means ± SD of three independent experiments. * p < 0.05, ** p < 0.01, compared to the LPS-treated group.
Figure 4
Figure 4
Effects of HB on LPS-induced production of IL-6 (a) and IL-1β (b) in RAW 264.7 Cells. RAW 264.7 Cells were pretreated with different concentrations of HB (25, 50, and 100 μM) for 1 h before treatment with 1 μg/mL LPS. After incubation for 18 h, the IL-6 and IL-1β production analysis was detected by ELISA kits. The values are presented as means ± SD of three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, compared to the LPS-treated group.
Figure 5
Figure 5
Effects of HB on LPS-induced ROS production in RAW 264.7 Cells measured by flow cytometry. (A) Control (B) LPS (1 μg/mL) (C) RAW 264.7 Cells were pretreated with 100 μM HB for 1 h before treatment with 1 μg/mL LPS for 18 h. (D) HB reduced the LPS-induced ROS production in RAW 264.7 Cells. Cells incubated with 1μg/mL LPS for 19 h were used as a positive controls. The values are presented as means ± SD of three independent experiments * p < 0.05, compared to the LPS-treated group.
Figure 5
Figure 5
Effects of HB on LPS-induced ROS production in RAW 264.7 Cells measured by flow cytometry. (A) Control (B) LPS (1 μg/mL) (C) RAW 264.7 Cells were pretreated with 100 μM HB for 1 h before treatment with 1 μg/mL LPS for 18 h. (D) HB reduced the LPS-induced ROS production in RAW 264.7 Cells. Cells incubated with 1μg/mL LPS for 19 h were used as a positive controls. The values are presented as means ± SD of three independent experiments * p < 0.05, compared to the LPS-treated group.
Figure 6
Figure 6
Effects of HB on LPS-induced apoptosis in RAW 264.7 Cells measured by flow cytometry. (A) Control (B) LPS (1 μg/mL) (C) RAW 264.7 Cells were pretreated with 100 μM HB for 1 h before treatment with 1 μg/mL LPS for 18 h. (D) HB reduced the LPS-induced apoptosis in RAW 264.7 Cells. Cells incubated with 1 μg/mL LPS for 19 h were used as a positive controls. The values are presented as means ± SD of three independent experiments * p < 0.05, compared to the LPS-treated group.
Figure 7
Figure 7
Effect of HB on LPS-induced expression of IL-6 and IL-1β in RAW 264.7 cells. RAW 264.7 cells were pretreated with 50 and 100 μM HB for 1 h before treatment with 1 μg/mL LPS. After incubation for 18 h, the expression of levels of IL-6 and IL-1β mRNA were measured by RT-PCR analysis. The data is expressed as mean folds of the mRNA expression versus LPS-stimulated group. The values are presented as means ± SD of three independent experiments. * p < 0.05, compared to the LPS-treated group.
Figure 8
Figure 8
Effects of HB on LPS-induced PI3K and AKT phosphorylation. RAW 264.7 cells were pretreated with 50 and 100 μM HB for 1 h before treatment with 1 μg/mL LPS. After incubation for 30 min, the expression of levels of PI3K and AKT protein were measured by western blot analysis. The values are presented as means ± SD of three independent experiments. * p < 0.05, compared to the LPS-treated group.
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References

    1. Cheng B.C.Y., Ma X.Q., Kwan H.Y., Tse K.W., Cao H.H., Su T., Shu X., Wu Z.Z., Yu Z.L. A herbal formula consisting of Rosae Multiflorae Fructus and Lonicerae Japonicae Flos inhibits inflammatory mediators in LPS-stimulated RAW 264.7 macrophages. J. Ethnopharmacol. 2014;153:922–927. doi: 10.1016/j.jep.2014.02.029. - DOI - PubMed
    1. Wang J., Mazza G. Inhibitory effects of anthocyanins and other phenolic compounds on nitric oxide production in LPS/IFN-γ-activated RAW 264.7 macrophages. J. Agric. Food Chem. 2002;50:850–857. doi: 10.1021/jf010976a. - DOI - PubMed
    1. Kundu J.K., Surh Y.J. Inflammation: Gearing the journey to cancer. Mutat. Res. 2008;659:15–30. doi: 10.1016/j.mrrev.2008.03.002. - DOI - PubMed
    1. Ning-Hua T., Jun Z., Chang-Xiang C., Shou-Xun Z. Cyclopeptides from the roots of Pseudostellaria heterophylla. Phytochemistry. 1993;32:1327–1330. doi: 10.1016/S0031-9422(00)95114-8. - DOI
    1. Tan N.H., Zhou J. Plant cyclopeptides. Chem. Rev. 2006;106:840–895. doi: 10.1021/cr040699h. - DOI - PubMed

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