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. 2022 Jun 21:2022:7767940.
doi: 10.1155/2022/7767940. eCollection 2022.

Transcriptomic Profile Analysis of Streptococcus mutans Response to Acmella paniculata Flower Extracts

Siti Aisyah Abd Ghafar  1 , Nur Syahirah Salehuddin  1 , Nor Zaihana Abdul Rahman  1 , Nadia Halib  1 , Rohazila Mohamad Hanafiah  1
Affiliations

Transcriptomic Profile Analysis of Streptococcus mutans Response to Acmella paniculata Flower Extracts

Siti Aisyah Abd Ghafar et al. Evid Based Complement Alternat Med. .

Abstract

Background: Acmella paniculata has been used as a traditional medicine to treat oral health diseases such as dental caries and periodontitis. Streptococcus mutans is a common bacterium that initiates dental caries at an early stage.

Aim: The aim of this study was to determine the mode of action of A. paniculata (extracts) against S. mutans growth.

Methods: Time-kill assay has been done to investigate the rate of kill and effectiveness of Acmella paniculata (AP) extracts against S. mutans growth. Phytochemical analysis was done to identify major compounds in AP extracts using gas chromatography mass spectrometry (GCMS). Scanning and transmission electron microscopy (SEM and TEM) have been done to observe the morphological changes of treated bacteria. Transcriptomic profile analysis has been done using Next Gene Sequencing.

Results: AP flower n-hexane (APFH) and AP flower dichloromethane (APFD) extracts acted as bactericidal agents after killing >3 log10 cfu/mL of S. mutans after 24 hours. Oleic and hexadecenoic acids were found to be the major compounds in APFD and APFH extracts, respectively. Photomicrographs from SEM and TEM of treated S. mutans show that the bacterial cell wall has been lysed and the cytoplasm content was decreased. Pathway analysis revealed that the APFD extract significantly affected biosynthesis peptidoglycan, gene expression, RNA processing, and macromolecule metabolism processes in S. mutans.

Conclusion: Data analysis revealed that multiple mechanisms of action were involved in antibacterial activity of A. paniculata extracts toward S. mutans.

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Conflict of interest statement

The authors declare that they have no conflicts of interest.

Figures

Figure 1
Figure 1
SEM analysis of S. mutans treated with APFH and APFD compared with untreated cells: (a) untreated S. mutans, (b) S. mutans treated with APFH (12.5 mg/mL), (c) S. mutans treated with APFH (50 mg/mL), (d) S. mutans treated with APFD (12.5 mg/mL), and (e) S. mutans treated with APFD (50 mg/mL). The morphology of untreated cell was aggregated (i); meanwhile, treated cells were elongated (ii), shrunk (iii), small (iv), and lysed (v).
Figure 2
Figure 2
TEM analysis of S. mutans treated with APFH and APFD compared with untreated cells: (a) untreated S. mutans, (b) S. mutans treated with APFH (12.5 mg/mL), (c) S. mutans treated with APFH (50 mg/mL), (d) S. mutans treated with APFD (12.5 mg/mL), and (e) S. mutans treated with APFD (50 mg/mL). Some characteristics of untreated bacteria were clearly seen such as granules, vacuoles and encapsulated structure (i); meanwhile, the cell wall of treated bacteria was lysed (ii) and distorted (iii) with formation of hollows (iv).
Figure 3
Figure 3
Total ion chromatogram of APFD extract. The highest peak for the APFD extract was linoelaidic acid (retention time: 39.46).
Figure 4
Figure 4
Total ion chromatogram of the APFH extract. The highest peak for the APFH extract was hexadecanoic acid (retention time: 34.63).
Figure 5
Figure 5
Total DEGs of S. mutans after treatment with APFD extract. Out of 1266 genes, 622 were downregulated and 644 were upregulated.
Figure 6
Figure 6
DEG analysis of S. mutans treated with APFD (MIC). (a) Heatmap clustering of all DEGs (1 < log2FC > 1 and p value < 0.05) in S. mutans treated with APFD. Each row represents one sample, and each column represents one gene. (b) Volcano image of gene regulation of S. mutans treated with APFD (MIC). Green color represents downregulated genes, and red color represents upregulated genes.
Figure 7
Figure 7
(a) Gene ontology category of DEGs in molecular function. The highest downregulated gene in molecular function was nucleic acid binding. (b) Gene ontology category of DEGs in cellular component (CC). Genes of cytoplasm were the highest downregulated genes in cellular component. (c) Gene ontology category of biology process (BP) showing downregulated genes in respective processes.
Figure 8
Figure 8
KEGG enrichment scatter plot of DEGs (downregulated). The y-axis shows the name of the pathway and the x-axis shows the rich factor. Dot size represents the number of different genes, and the color indicates the q value.
See this image and copyright information in PMC

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