Cepharanthine ameliorates dextran sulphate sodium-induced colitis through modulating gut microbiota

Abstract

Cepharanthine (CEP) is an active alkaloid isolated from Stephania Cepharantha Hayata. It is reported that the anti-inflammatory properties of CEP could be employed to treat a variety of diseases. In this study, we first found that CEP ameliorates ulcerative colitis (UC) induced by DSS. The effect of CEP on gut microbiota was further evaluated by 16S rRNA gene sequencing, antibiotic pretreatment and faecal microbiota transplantation (FMT). Results showed that the abundances of gut microbiota, such as Romboutsia, Turicibacter and Escherichia-Shigella (especially Romboutsia), were significantly reduced after CEP treatment. Additionally, we explored the mechanisms of CEP by a strategy integrating transcriptomics with network pharmacology. The transcriptome data confirmed that CEP functioned through cytokine and cytokine receptor pathways. The expression levels of 10 pro-inflammatory hub genes (such as CXCL1, CXCL9, CCL7) were positively correlated with the abundance of Romboutsia. Our data identified Romboutsia as a potential pathobiont in UC. Collectively, we confirmed that CEP relieved colon inflammation by modulating gut microbiota and pro-inflammatory cytokine expression. CEP can be adopted to design novel effective therapeutic strategies for UC.

Fig. 1

CEP treatment ameliorates DSS‐induced colitis in Mice. (A) the percentage of the weight of mice was measured for 7 days; (B)The DAI score; (C) Photographs of the colon; (D) The colon length; (E) H&E staining for colon tissues. (100×; 200×) (F) The Histology score; Values are presented as the mean ± standard error of the mean (SEM); CON group, n = 7; DSS group, n = 7; DSS + CEP group, n = 4; *P < 0.05, **P < 0.01, ***P < 0.001.

Fig. 2

Cepharanthine regulates the structure and composition of gut microbiota. (A)The Venn diagram of CON, DSS, and DSS + CEP group; (B) PCoA score (C) Chao index (D) Shannon index; (E) Bar chart of gut microbiota at the phylum level; (F) Heatmap of gut microbiota at the genus level; (G) Relative abundance of discriminative gut microbiota at the genus level; (H) The LEfSe analysis; (I) The metastats analysis between DSS and DSS + CEP group; Values are presented as the mean ± standard error of the mean (SEM); CON group, n = 7; DSS group, n = 7; DSS + CEP group, n = 4; *P < 0.05, **P < 0.01, ***P < 0.001.

Fig. 3

FMT treatment ameliorates intestinal inflammation by regulating the gut microbiota. (A) the percentage of the weight of mice was measured for 7 days; (B)The DAI score; (C) Photographs of the colon; (D) The colon length; (E) H&E staining for colon tissues. (100×; 200×) (F) The Histology score; Values are presented as the mean ± standard error of the mean (SEM); DSS group, n = 4; DSS + FMT group, n = 5; *P < 0.05, **P < 0.01, ***P < 0.001.

Fig. 4

(A) Heatmap of gut microbiota at the phylum level; (B) Heatmap of gut microbiota at the genus level; (C) Relative abundance of discriminative gut microbiota at the genus level; *P < 0.05.

Fig. 5

Effects of Abx pretreatment on the CEP‐treated colitis. (A) Treatment diagram of Abx + DSS and Abx + DSS + CEP group; (B) the percentage of the weight of mice was measured for 7 days; (C)The DAI score; The colon length; (D) Photographs of colon; (E) H&E staining for colon tissues. (100×; 200×) (F) The Histology score; Values are presented as the mean ± standard error of the mean (SEM); n = 6 for each treatment. *P < 0.05, **P < 0.01, ***P < 0.001.

Fig. 6

CEP affects Cytokine‐cytokine receptor interaction pathways. (A) Volcano plot analysis identifies DEGs between DSS and DSS + CEP group; (B) Go term enrichment analysis of DEGs; (C) KEGG pathway analysis of DEGs. Correlation analysis between gut microbiota and significant genus. (D) String software analyzed the protein‐protein network; (E) The most significant module identified by MCODE (score = 17.059). (F) The spearman's correlation analysis between the DEGs and Romboutsia genus by SPSS software.