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. 2018 Apr 16:9:371.
doi: 10.3389/fphar.2018.00371. eCollection 2018.

Inhibition of Streptococcus suis Adhesion and Biofilm Formation in Vitro by Water Extracts of Rhizoma Coptidis

Yan-Hua Li  1   2 , Yong-Hui Zhou  1   2 , Yong-Zhi Ren  1   2 , Chang-Geng Xu  1   2 , Xin Liu  1   2 , Bing Liu  1   2 , Jian-Qing Chen  1   2 , Wen-Ya Ding  1   2 , Yu-Lin Zhao  1   2 , Yan-Bei Yang  1   2 , Shuai Wang  1   2 , Di Liu  3
Affiliations

Inhibition of Streptococcus suis Adhesion and Biofilm Formation in Vitro by Water Extracts of Rhizoma Coptidis

Yan-Hua Li et al. Front Pharmacol. .

Abstract

Streptococcus suis is difficult to treat and responsible for various infections in humans and pigs. It can also form biofilms and induce persistent infections. Rhizoma Coptidis is a medicinal plant widely used in Traditional Chinese Medicine. Although the inhibitory effects of Rhizoma Coptidis on biofilm formation have been investigated in several studies, the ability of Rhizoma Coptidis to inhibit S. suis biofilm formation and the underlying mechanisms have not yet been reported. In this study, we showed that sub-minimal inhibitory concentrations (25 and 50 μg mL-1) of water extracts of Rhizoma Coptidis (Coptis deltoidea C.Y.Cheng & P.K.Hsiao, obtained from Sichuan Province) were sufficient to inhibit biofilm formation, as shown in the tissue culture plate (TCP) method and scanning electron microscopy. Real-time PCR and iTRAQ were used to measure gene and protein expression in S. suis. Sub-minimum inhibitory concentrations (25 and 50 μg mL-1) of Rhizoma Coptidis water extracts inhibited S. suis adhesion significantly in an anti-adherence assay. Some genes, such as gapdh, sly, and mrp, and proteins, such as antigen-like protein, CPS16V, and methyltransferase H, involved in adhesion were significantly modulated in cells treated with 50 μg mL-1 of Rhizoma Coptidis water extracts compared to untreated cells. The results from this study suggest that compounds in Rhizoma Coptidis water extracts play an important role in inhibiting adhesion of S. suis cells and, therefore, biofilm formation.

Keywords: Rhizoma Coptidis; S. suis; adhesion; biofilm; iTRAQ technology; real-time PCR.

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Figures

FIGURE 1
FIGURE 1
The HPLC chromatograms of the Rhizoma Coptidis aqueous extract (black, retention time 15.43 min) and the berberine standard (red, retention time 15.43 min).
FIGURE 2
FIGURE 2
Effect of Rhizoma Coptidis at different concentrations on S. suis ATCC700794 biofilm formation. Data are expressed as means ± standard. Significant decrease (∗p<0.05) compared to control biofilm formation of S. suis in vitro.
FIGURE 3
FIGURE 3
Scanning electron microscopy of biofilm of S. suis grown in THB broth. (A) With 0 MIC (0 μg mL-1) concentration of aqueous extracts of Rhizoma Coptidis; (B) with 1/2 × MIC (50 μg mL-1) concentration of aqueous extracts of Rhizoma Coptidis.
FIGURE 4
FIGURE 4
Effect of Rhizoma Coptidis at different concentrations on S. suis ATCC700794 adhesion to glass (A), or PK-15 cells (B). Data are expressed as means ± standard. Significant decrease (∗p < 0.05) compared to control in vitro.
FIGURE 5
FIGURE 5
Effect of 1/2MIC of aqueous extracts of Rhizoma Coptidis on mRNA decreased expression of genes in S. suis ATCC700794. Data are expressed as means ± standard deviations. The expression was normalized to 16S rRNA. Controls refer to the absence of aqueous extracts of Rhizoma Coptidis. Significantly different (∗p < 0.05) compared to untreated control bacteria.
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