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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

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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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Effect of red ginseng acidic polysaccharide (RGAP) on invasion and intracellular growth of Brucella (B.) abortus. (A) Bacterial internalization efficiency. (B) Bacterial intracellular growth efficiency. Data represent the means ± SD of duplicate samples in at least three independent experiments. Statistically significant differences relative to the untreated control are indicated by asterisks (*p < 0.05; **p < 0.01; ***p < 0.001).
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Figure 1: Effect of red ginseng acidic polysaccharide (RGAP) on invasion and intracellular growth of Brucella (B.) abortus. (A) Bacterial internalization efficiency. (B) Bacterial intracellular growth efficiency. Data represent the means ± SD of duplicate samples in at least three independent experiments. Statistically significant differences relative to the untreated control are indicated by asterisks (*p < 0.05; **p < 0.01; ***p < 0.001).

Mentions: The invasion of B. abortus in RAW 264.7 cells was verified by pre-incubation with the highest non-cytotoxic concentration of RGAP (0.1 mg/mL) for the indicated times (0, 30 min). The results indicate that the invasion of B. abortus into RGAP-treated cells was significantly reduced compared with the untreated control (p < 0.05) (panel A in Fig. 1). Furthermore, RGAP-treated cells displayed significantly reduced intracellular growth of B. abortus at 2, 24 and 48 h post-infection compared with the untreated control cells (panel B in Fig. 1).


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 red ginseng acidic polysaccharide (RGAP) on invasion and intracellular growth of Brucella (B.) abortus. (A) Bacterial internalization efficiency. (B) Bacterial intracellular growth efficiency. Data represent the means ± SD of duplicate samples in at least three independent experiments. Statistically significant differences relative to the untreated control are indicated by asterisks (*p < 0.05; **p < 0.01; ***p < 0.001).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 1: Effect of red ginseng acidic polysaccharide (RGAP) on invasion and intracellular growth of Brucella (B.) abortus. (A) Bacterial internalization efficiency. (B) Bacterial intracellular growth efficiency. Data represent the means ± SD of duplicate samples in at least three independent experiments. Statistically significant differences relative to the untreated control are indicated by asterisks (*p < 0.05; **p < 0.01; ***p < 0.001).
Mentions: The invasion of B. abortus in RAW 264.7 cells was verified by pre-incubation with the highest non-cytotoxic concentration of RGAP (0.1 mg/mL) for the indicated times (0, 30 min). The results indicate that the invasion of B. abortus into RGAP-treated cells was significantly reduced compared with the untreated control (p < 0.05) (panel A in Fig. 1). Furthermore, RGAP-treated cells displayed significantly reduced intracellular growth of B. abortus at 2, 24 and 48 h post-infection compared with the untreated control cells (panel B in Fig. 1).

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