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. 2023 Mar 2;24(5):4844.
doi: 10.3390/ijms24054844.

Study on Anti-Constipation Effects of Hemerocallis citrina Baroni through a Novel Strategy of Network Pharmacology Screening

Yuxuan Liang  1 , Xiaoyi Wei  1 , Rui Ren  1 , Xuebin Zhang  1 , Xiyao Tang  1 , Jinglan Yang  1 , Xiaoqun Wei  1 , Riming Huang  1 , Gary Hardiman  2 , Yuanming Sun  1   3 , Hong Wang  1   3
Affiliations

Study on Anti-Constipation Effects of Hemerocallis citrina Baroni through a Novel Strategy of Network Pharmacology Screening

Yuxuan Liang et al. Int J Mol Sci. .

Abstract

Daylily (Hemerocallis citrina Baroni) is an edible plant widely distributed worldwide, especially in Asia. It has traditionally been considered a potential anti-constipation vegetable. This study aimed to investigate the anti-constipation effects of daylily from the perspective of gastro-intestinal transit, defecation parameters, short-chain organic acids, gut microbiome, transcriptomes and network pharmacology. The results show that dried daylily (DHC) intake accelerated the defecation frequency of mice, while it did not significantly alter the levels of short-chain organic acids in the cecum. The 16S rRNA sequencing showed that DHC elevated the abundance of Akkermansia, Bifidobacterium and Flavonifractor, while it reduced the level of pathogens (such as Helicobacter and Vibrio). Furthermore, a transcriptomics analysis revealed 736 differentially expressed genes (DEGs) after DHC treatment, which are mainly enriched in the olfactory transduction pathway. The integration of transcriptomes and network pharmacology revealed seven overlapping targets (Alb, Drd2, Igf2, Pon1, Tshr, Mc2r and Nalcn). A qPCR analysis further showed that DHC reduced the expression of Alb, Pon1 and Cnr1 in the colon of constipated mice. Our findings provide a novel insight into the anti-constipation effects of DHC.

Keywords: 16S rRNA; constipation; daylily; network pharmacology; transcriptomes.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Gastrointestinal transit rate and defecation frequency in six hours of mice. Normal: distilled water-treated; Lop: Loperamide-treated; DHC: dry daylily-treated. (a) Gastrointestinal transit rate, (b) fecal number in six hours on day 13 and (c) fecal number in six hours on day 14; # represents the comparison with Lop group, p < 0.05; ## represents the comparison with Lop group, p < 0.01; * represents the comparison with Normal group, p < 0.05; ** represents the comparison with Normal group, p < 0.01.
Figure 2
Figure 2
Short-chain organic acid content. Normal: distilled water-treated; Lop: Loperamide-treated; DHC: dry daylily-treated. (a–f) acetic acid, propionic acid, butyric acid, valeric acid, isobutyric acid and isovaleric acid.
Figure 3
Figure 3
Alpha diversity and Beta diversity analysis. Normal: distilled water-treated; Lop: Loperamide-treated; DHC: dry daylily-treated. (a) Alpha diversity (Observed otus, Shannon and Chao1) and (b) Beta diversity (Principal component analysis).
Figure 4
Figure 4
Intestinal flora composition analysis. Normal: distilled water-treated; Lop: Loperamide-treated; DHC: dry daylily-treated. (a) Relative abundance of the top 30 genera with a heatmap, (b) genera that changed significantly were between Lop and Normal and (c) genera that changed significantly were between DHC and Lop.
Figure 5
Figure 5
Transcriptomic analysis. (a) differential expression genes in different groups, (b) volcano plot of differential expression genes between Lop and Normal, (c) volcano plot of differential expression genes between DHC and Lop, (d) KEGG pathway enrichment of differential expression genes between Lop and Normal and (e) KEGG pathway enrichment of differential expression genes between DHC and Lop.
Figure 6
Figure 6
Joint analysis of network pharmacology and transcriptomes. (a) Venn diagram for the differential expression genes (Lop vs. Normal) and constipation targets and (b) Protein–protein interaction network of overlapping targets.
Figure 7
Figure 7
Relative expression of genes (mean ± SD). Normal: distilled water-treated; Lop: Loperamide-treated; DHC: dry daylily-treated; # represents the comparison with Lop group, p < 0.05.
See this image and copyright information in PMC

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