Sodium houttuyfonate effectively treats acute pulmonary infection of Pseudomonas aeruginosa by affecting immunity and intestinal flora in mice.

Introduction: Pseudomonas aeruginosa is a major nosocomial pathogen that frequently causes ventilator-associated pneumonia in specific populations. Sodium houttuyfonate (SH) has shown mild antibacterial activity against P. aeruginosa in vitro, but the mechanism of potent antimicrobial activity of SH against P. aeruginosa infection in vivo remains unclear. Methods: Here, using the mouse pneumonia model induced by P. aeruginosa nasal drip to explore the therapeutic effects of SH. Results: We found that SH exhibits dose-dependent therapeutic effects of reducing P. aeruginosa burden and systemic inflammation in pneumonia mice. SH ameliorates inflammatory gene expression and production of inflammatory proteins, such as interleukin-6 (IL-6), nuclear factor kappa-B (NF-κB) and toll-like receptor 4 (TLR4), associated with the TLR4/NF-κB pathway in mice with P. aeruginosa pneumonia. Furthermore, we analyzed the intestinal flora of mice and found that compared with the model group, the abundance and diversity of beneficial bacterial flora of SH treatment groups increased significantly, suggesting that SH can improve the intestinal flora disorder caused by inflammation. In addition, SH improves alpha and beta diversity index and reduces species abundance differences of intestinal flora in pneumonia mice. Discussion: Taken together, our presented results indicate that SH may effectively alleviate the acute pulmonary infection induced by P. aeruginosa by reducing the disturbance of regulating immunity and intestinal flora in mice.

Sodium New Houttuyfonate Affects Transcriptome and Virulence Factors of Pseudomonas aeruginosa Controlled by Quorum Sensing.

As a major opportunistic pathogen, Pseudomonas aeruginosa can produce various virulence factors and form biofilms. These processes are controlled by the quorum sensing (QS) system. Sodium new houttuyfonate (SNH) is an adduct of houttuyfonate, the main component of the common Chinese medicine plant Houttuynia cordata, which has antibacterial and anti-inflammatory effects. We evaluated the effect of SNH on P. aeruginosa biofilms, virulence factors, and transcription. Transcriptome analysis showed that the key rhlI and pqsA genes of the P. aeruginosa QS system were down-regulated after SNH treatment. SNH reduces proteases and pyocyanin production and inhibits biofilm formation by regulating the P. aeruginosa QS system. SNH also changes the expression of genes related to virulence factors and biofilms (lasA, lasB, lecA, phzM, pqsA, and pilG). These results suggested that the mechanism of SNH against P. aeruginosa by affecting the expression of biofilm and virulence factors controlled by quorum sensing.

Sodium New Houttuyfonate Inhibits Candida albicans Biofilm Formation by Inhibiting the Ras1-cAMP-Efg1 Pathway Revealed by RNA-seq.

Here, we aim to investigate the antifungal effect and mechanism of action of sodium new houttuyfonate (SNH) against Candida albicans. Microdilution analysis results showed that SNH possesses potent inhibitory activity against C. albicans SC5314, with a MIC80 of 256 µg/mL. Furthermore, we found that SNH can effectively inhibit the initial adhesion of C. albicans. Inverted microscopy, crystal violet staining, scanning electron microscopy and confocal laser scanning microscopy results showed that morphological changes during the transition from yeast to hypha and the biofilm formation of C. albicans are repressed by SNH treatment. We also found that SNH can effectively inhibit the biofilm formation of clinical C. albicans strains (Z103, Z3044, Z1402, and Z1407) and SNH in combination with fluconazole, berberine chloride, caspofungin and itraconazole antifungal agents can synergistically inhibit the biofilm formation of C. albicans. Eukaryotic transcriptome sequencing and qRT-PCR results showed that SNH treatment resulted in significantly down-regulated expression in several biofilm formation related genes in the Ras1-cAMP-Efg1 pathway (ALS1, ALA1, ALS3, EAP1, RAS1, EFG1, HWP1, and TEC1) and significantly up-regulated expression in yeast form-associated genes (YWP1 and RHD1). We also found that SNH can effectively reduce the production of key messenger cAMP in the Ras1-cAMP-Efg1 pathway. Furthermore, using Galleria mellonella as an in vivo model we found that SNH can effectively treat C. albicans infection in vivo. Our presented results suggest that SNH exhibits potential antibiofilm effects related to inhibiting the Ras1-cAMP-Efg1 pathway in the biofilm formation of C. albicans.

Effects of sodium houttuyfonate on transcriptome of Pseudomonas aeruginosa.

OBJECTIVES: The purpose of this experiment is to analyze the changes of transcriptome in Pseudomonas aeruginosa under the action of sodium houttuyfonate (SH) to reveal the possible mechanism of SH inhibiting P. aeruginosa. We analyzed these data in order to compare the transcriptomic differences of P. aeruginosa in SH treatment and blank control groups. DATA DESCRIPTION: In this project, RNA-seq of BGISEQ-500 platform was used to sequence the transcriptome of P. aeruginosa, and sequencing data of 8 samples of P. aeruginosa are generated as follows: SH treatment (SH1, SH2, SH3, SH4), negative control (Control 1, Control 2, Control 3, Control 4). Quality control is carried out on raw reads to determine whether the sequencing data is suitable for subsequent analysis. Totally 170.53 MB of transcriptome sequencing data is obtained. Then the filtered clean reads are aligned and compared to the reference genome to proceed second quality control. After completion, 5938 genes are assembled from sequencing data. Further quantitative analysis of genes and screening of differentially expressed genes based on gene expression level reveals that there are 2047 significantly differentially expressed genes under SH treatment, including 368 up-regulated genes and 1679 down-regulated genes.