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Inhibitory effect of red ginseng acidic polysaccharide from Korean red ginseng on phagocytic activity and intracellular replication of Brucella abortus in RAW 264.7 cells

View Article: PubMed Central - PubMed

ABSTRACT

Korean red ginseng (KRG) has long been used in traditional Korean and Oriental medicine. However, the anti-bacterial mechanism and therapeutic efficiency of KGR for intracellular Brucella infection are still unclear. In this study, the bactericidal activity of Korean red ginseng acidic polysaccharide (RGAP) on Brucella (B.) abortus and its cytotoxic effects on RAW 264.7 cells were evaluated. In addition, B. abortus internalization and intracellular replication in macrophages were investigated after RGAP treatment. RGAP-incubated cells displayed a marked reduction in the adherence, internalization and intracellular growth of B. abortus in macrophages. Furthermore, decreased F-actin fluorescence was observed relative to untreated B. abortus-infected cells. Western blot analysis of intracellular signaling proteins revealed reduced ERK, JNK and p38α phosphorylation levels in B. abortus-infected RGAP-treated cells compared to the control. Moreover, elevated co-localization of B. abortus-containing phagosomes with lysosome-associated membrane protein 1 (LAMP-1) were observed in RGAP-treated cells compared with the control. Overall, the results of this study suggest that RGAP can disrupt phagocytic activity of B. abortus via suppression of mitogen-activated protein kinases (MAPKs) signaling proteins ERK, JNK and p38 levels and inhibit intracellular replication of B. abortus by enhancing phagolysosome fusion, which may provide an alternative control of brucellosis.

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Fluorescence-activated cell sorting (FACS) analysis of the effects of RGAP on phagocytosis of B. abortus by F-actin polymerization modulation. (A) F-actin polymerization and bacterial co-localization (scale bars = 5 µm). (B) FACS analysis for F-actin content. (C) Intensification of F-actin polymerization. The data shown are representative of at least three independent experiments. Statistically significant differences relative to untreated control cells are indicated by an asterisk (*p < 0.05). DIC, differential interference contrast.
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Figure 3: Fluorescence-activated cell sorting (FACS) analysis of the effects of RGAP on phagocytosis of B. abortus by F-actin polymerization modulation. (A) F-actin polymerization and bacterial co-localization (scale bars = 5 µm). (B) FACS analysis for F-actin content. (C) Intensification of F-actin polymerization. The data shown are representative of at least three independent experiments. Statistically significant differences relative to untreated control cells are indicated by an asterisk (*p < 0.05). DIC, differential interference contrast.

Mentions: Phalloidin-associated F-actin fluorescence microscopy indicated diminished F-actin polymerization for B. abortus invasion in RGAP-treated cells compared with untreated control cells and showed a reduction in filopodia and lamellipodia scattering in the peripheral cells (panel A in Fig. 3). Because F-actin polymerization is required for B. abortus phagocytosis, the F-actin content of RGAP-treated cells upon B. abortus invasion was assessed and quantified. Fluorescence-activated cell sorting (FACS) analysis revealed that treatment of murine macrophages with RGAP significantly reduced the F-actin content in these cells upon bacterial infection, as indicated by decreased F-actin fluorescence intensity compared with B. abortus-infected untreated cells (panel B and C in Fig. 3). The F-actin content of the uninfected RGAP-treated cells showed no significant difference from the untreated control cells.


Inhibitory effect of red ginseng acidic polysaccharide from Korean red ginseng on phagocytic activity and intracellular replication of Brucella abortus in RAW 264.7 cells
Fluorescence-activated cell sorting (FACS) analysis of the effects of RGAP on phagocytosis of B. abortus by F-actin polymerization modulation. (A) F-actin polymerization and bacterial co-localization (scale bars = 5 µm). (B) FACS analysis for F-actin content. (C) Intensification of F-actin polymerization. The data shown are representative of at least three independent experiments. Statistically significant differences relative to untreated control cells are indicated by an asterisk (*p < 0.05). DIC, differential interference contrast.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5037298&req=5

Figure 3: Fluorescence-activated cell sorting (FACS) analysis of the effects of RGAP on phagocytosis of B. abortus by F-actin polymerization modulation. (A) F-actin polymerization and bacterial co-localization (scale bars = 5 µm). (B) FACS analysis for F-actin content. (C) Intensification of F-actin polymerization. The data shown are representative of at least three independent experiments. Statistically significant differences relative to untreated control cells are indicated by an asterisk (*p < 0.05). DIC, differential interference contrast.
Mentions: Phalloidin-associated F-actin fluorescence microscopy indicated diminished F-actin polymerization for B. abortus invasion in RGAP-treated cells compared with untreated control cells and showed a reduction in filopodia and lamellipodia scattering in the peripheral cells (panel A in Fig. 3). Because F-actin polymerization is required for B. abortus phagocytosis, the F-actin content of RGAP-treated cells upon B. abortus invasion was assessed and quantified. Fluorescence-activated cell sorting (FACS) analysis revealed that treatment of murine macrophages with RGAP significantly reduced the F-actin content in these cells upon bacterial infection, as indicated by decreased F-actin fluorescence intensity compared with B. abortus-infected untreated cells (panel B and C in Fig. 3). The F-actin content of the uninfected RGAP-treated cells showed no significant difference from the untreated control cells.

View Article: PubMed Central - PubMed

ABSTRACT

Korean red ginseng (KRG) has long been used in traditional Korean and Oriental medicine. However, the anti-bacterial mechanism and therapeutic efficiency of KGR for intracellular Brucella infection are still unclear. In this study, the bactericidal activity of Korean red ginseng acidic polysaccharide (RGAP) on Brucella (B.) abortus and its cytotoxic effects on RAW 264.7 cells were evaluated. In addition, B. abortus internalization and intracellular replication in macrophages were investigated after RGAP treatment. RGAP-incubated cells displayed a marked reduction in the adherence, internalization and intracellular growth of B. abortus in macrophages. Furthermore, decreased F-actin fluorescence was observed relative to untreated B. abortus-infected cells. Western blot analysis of intracellular signaling proteins revealed reduced ERK, JNK and p38&alpha; phosphorylation levels in B. abortus-infected RGAP-treated cells compared to the control. Moreover, elevated co-localization of B. abortus-containing phagosomes with lysosome-associated membrane protein 1 (LAMP-1) were observed in RGAP-treated cells compared with the control. Overall, the results of this study suggest that RGAP can disrupt phagocytic activity of B. abortus via suppression of mitogen-activated protein kinases (MAPKs) signaling proteins ERK, JNK and p38 levels and inhibit intracellular replication of B. abortus by enhancing phagolysosome fusion, which may provide an alternative control of brucellosis.

No MeSH data available.


Related in: MedlinePlus