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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.

No MeSH data available.


Effect of RGAP on activation of intracellular signaling for the phagocytosis of B. abortus. Immunoblot analyses of total RAW 264.7 cell lysates pre-treated with RGAP were assessed using phospho-specific ERK1/2, JNK and p38α antibodies at the indicated times. Images shown are representative of at least three independent experiments.
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Figure 4: Effect of RGAP on activation of intracellular signaling for the phagocytosis of B. abortus. Immunoblot analyses of total RAW 264.7 cell lysates pre-treated with RGAP were assessed using phospho-specific ERK1/2, JNK and p38α antibodies at the indicated times. Images shown are representative of at least three independent experiments.

Mentions: The phosphorylation levels of ERK1/2, JNK and p38α in RGAP-treated cells at 30 min post-infection were reduced by 27.58%, 34.76% and 47.92%, respectively, compared with B. abortus-infected control cells (Fig. 4). These findings indicate that RGAP suppressed the activation of MAPKs in RAW 264.7 cells, which inhibited the invasion of B. abortus into macrophages. In addition, co-localization of LAMP-1 with BCPs in RGAP-treated cells was observed to markedly increase, showing a significant increase of up to 2.03-fold after 2 h of incubation relative to untreated control cells (p < 0.001) (Fig. 5).


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
Effect of RGAP on activation of intracellular signaling for the phagocytosis of B. abortus. Immunoblot analyses of total RAW 264.7 cell lysates pre-treated with RGAP were assessed using phospho-specific ERK1/2, JNK and p38α antibodies at the indicated times. Images shown are representative of at least three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 4: Effect of RGAP on activation of intracellular signaling for the phagocytosis of B. abortus. Immunoblot analyses of total RAW 264.7 cell lysates pre-treated with RGAP were assessed using phospho-specific ERK1/2, JNK and p38α antibodies at the indicated times. Images shown are representative of at least three independent experiments.
Mentions: The phosphorylation levels of ERK1/2, JNK and p38α in RGAP-treated cells at 30 min post-infection were reduced by 27.58%, 34.76% and 47.92%, respectively, compared with B. abortus-infected control cells (Fig. 4). These findings indicate that RGAP suppressed the activation of MAPKs in RAW 264.7 cells, which inhibited the invasion of B. abortus into macrophages. In addition, co-localization of LAMP-1 with BCPs in RGAP-treated cells was observed to markedly increase, showing a significant increase of up to 2.03-fold after 2 h of incubation relative to untreated control cells (p < 0.001) (Fig. 5).

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.