AKT-targeted anti-inflammatory activity of Panax ginseng calyx ethanolic extract

Abstract

BackgroundKorean ginseng (Panax ginseng) plays an anti-inflammatory role in a variety of inflammatory diseases such as gastritis, hepatitis, and colitis. However, inflammation-regulatory activity of the calyx of the P. ginseng berry has not been thoroughly evaluated. To understand whether the calyx portion of the P. ginseng berry is able to ameliorate inflammatory processes, an ethanolic extract of P. ginseng berry calyx (Pg-C-EE) was prepared, and lipopolysaccharide-activated macrophages and HEK293 cells transfected with inflammation-regulatory proteins were used to test the anti-inflammatory action of Pg-C-EE.MethodsThe ginsenoside contents of Pg-C-EE were analyzed by HPLC. Suppressive activity of Pg-C-EE on NO production, inflammatory gene expression, transcriptional activation, and inflammation signaling events were examined using the Griess assay, reverse transcription-polymerization chain reaction, luciferase activity reporter gene assay, and immunoblotting analysis.ResultsPg-C-EE reduced NO production and diminished mRNA expression of inflammatory genes such as cyclooxygenase-2, inducible NO synthase, and tumor necrosis factor-α in a dose-dependent manner. This extract suppressed luciferase activity induced only by nuclear factor-κB. Interestingly, immunoblotting analysis results demonstrated that Pg-C-EE reduced the activities of protein kinase B (AKT)1 and AKT2.ConclusionThese results suggest that Pg-C-EE may have nuclear-factor-κB-targeted anti-inflammatory properties through suppression of AKT. The calyx of the P. ginseng berry is an underused part of the ginseng plant, and development of calyx-derived extracts may be useful for treatment of inflammatory diseases.

Regulation of 8-Hydroxydaidzein in IRF3-Mediated Gene Expression in LPS-Stimulated Murine Macrophages

“…As shown in Figure 3A, pre-treatment with 8-HD (50 µM) decreased the phosphorylation level of IRF-3 at Ser396. In similar conditions, we evaluated the level of IRF-3 translocated into the nucleus by using nuclear fractionation methods as previously reported [23]. Interestingly, Figure S2 indicated that 8-HD (50 µM) was also able to reduce IRF-3 translocation into the nucleus.…”

“…For this, 1 µg of total RNA was used for cDNA synthesis using a cDNA synthesis kit (Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturer's instructions. Analysis of mRNA expression by semiquantitative RT-PCR and qPCR was conducted as previously described [22,23]. The primer sequences used in this study are listed in Table 1.…”

Anti-Inflammatory Functions of Alverine via Targeting Src in the NF-κB Pathway

“…Next, 10 µL of MTT solution (5 mg/mL in phosphate-buffered saline, pH 7.4) was added to each well. After 3 h of incubation, 100 µL of MTT stop solution (15% SDS) was added to each well to solubilize the formazan, and the cells were incubated for 24 h, as reported previously [16]. The effects of alverine on cell viability were determined by measuring the absorbance at 570 nm using a SpectraMax 250 microplate reader.…”

In Vitro Effects of Dehydrotrametenolic Acid on Skin Barrier Function

“…Complementary DNA was synthesized from total RNA using MuLV reverse transcriptase, according to the manufacturer's instructions. Semi-quantitative RT-PCR and quantitative real-time PCR were conducted as previously described [15,44]. The primer sequences used in this study are listed in Table 1.…”

“…The epidermis maintains balance through the self-renewal of differentiated keratinocytes from the basal layer. The regeneration and repair of the skin are important for it to function properly [15]; thus, keratinocyte differentiation is essential for maintaining the skin barrier. Epidermis barrier formation is associated with various genes, such as involucrin, transglutaminase (TGM), and filaggrin (FLG) [16,17].…”