Article Title: Xianlian Jiedu Decoction alleviates colorectal cancer by regulating metabolic profiles, intestinal microbiota and metabolites

Journal: Phytomedicine

Impact Factor: 7.6

Affiliated Institution: School of Pharmacy, Nanjing University of Chinese Medicine

"With government support, publishing is not a problem." Traditional Chinese medicine (TCM) research has always been a hot topic. The typical research pattern is component identification → network pharmacology analysis → molecular docking analysis → experimental validation. These routines may have become tiresome. Today, I would like to share a different research strategy in TCM: a combination of non-targeted TCM analysis + gut microbiota + metabolomics. This approach is novel, highly reproducible, and you too can publish such a paper using this strategy.

Research Background

Colorectal cancer (CRC) is one of the most common malignant tumors in the world. With changes in dietary habits, CRC is increasingly affecting younger populations. Traditional Chinese medicine, due to its safety, low toxicity, and few side effects, has become a prominent alternative for prevention and treatment of CRC.

In addition, studies have shown that the development of CRC is closely associated with gut microbiota imbalance. Microbial-derived metabolites and short-chain fatty acids (SCFAs) may play key roles in preventing and treating CRC.

Xianlian Jiedu Decoction (XLJDD) has been used for decades in CRC treatment due to its remarkable therapeutic effects. Although its efficacy is widely recognized, its anti-CRC mechanisms remain unclear.

This study aims to clarify the inhibitory effects of XLJDD on CRC and the mechanisms by which it works via regulating metabolic characteristics, gut microbiota, and metabolites.

Research Strategy

First, non-targeted TCM analysis was used to identify the active components of XLJDD, followed by absolute quantification of key compounds.

Second, pharmacological efficacy experiments were performed on CRC model mice with different treatments:

· control group

· XLJDD low dose (11 g/kg/day)

· XLJDD high dose (44 g/kg/day)

· oxaliplatin group

Histopathology, immunofluorescence, and ELISA were then used to evaluate XLJDD's therapeutic effects against AOM/DSS-induced CRC.

Finally, mechanisms of action were investigated using:

· 16S rRNA sequencing

· SCFA quantification

· serum and fecal untargeted metabolomics

Chemical Composition Identification and Quantitative Analysis of XLJDD：

 

 

The main chemical components of XLJDD were identified, including alkaloids, organic acids, and flavonoids. Four key compounds—matrine, oxymatrine, palmatine, and berberine—were subjected to absolute quantitative analysis. This chemical profiling provides crucial information and quality assurance for further pharmacological research and clinical application of XLJDD.

XLJDD Inhibits Tumorigenesis in AOM/DSS-Induced Mice：

 

 

In the AOM/DSS mouse model, two different doses of XLJDD (11 and 44 g/kg/day) and oxaliplatin were administered. The impact on colorectal histology and tumor phenotype was examined.

Key observations:

· Oxaliplatin negatively impacted weight gain in mice;

· XLJDD significantly alleviated diarrhea, rectal prolapse, and weight loss;

· Survival curves showed that XLJDD improved survival rates in AOM/DSS-treated mice, while the oxaliplatin group showed a sharp decline in early-stage survival.

Compared to the model group:

· Both XLJDD and oxaliplatin groups showed significant reductions in tumor burden and tumor count;

· There was a notable increase in colon length and colon weight-to-length ratio.

These results suggest that XLJDD exhibits a dose-dependent anti-tumor effect in the AOM/DSS-induced CRC model.

XLJDD Mitigates Tissue Damage and Intestinal Inflammation

 

 

Hematoxylin and eosin (H&E) staining revealed that AOM/DSS treatment induced severe intestinal inflammation, marked by:

· Abnormal epithelial hyperplasia

· Increased nuclear division

· Significant inflammatory cell infiltration in colorectal tissue

However, both XLJDD and oxaliplatin groups showed:

· A reduction in mitotic activity

· Alleviation of inflammatory infiltration

Further analysis showed:

· β-catenin, a marker for colorectal precancerous lesions, was significantly reduced by XLJDD treatment.

· Expression levels of iNOS and COX-2, two pro-inflammatory enzymes associated with tumor risk, were elevated in the model group but significantly downregulated in both XLJDD and oxaliplatin groups.

Additionally, serum levels of pro-inflammatory cytokines (IL-6, IL-1β, TNF-α, IFN-γ) were markedly reduced after treatment. These results indicate that XLJDD can effectively suppress tumor-related inflammation and delay CRC progression.

XLJDD Promotes SCFA Release in AOM/DSS-Induced Mice：

 

 

The levels of six major short-chain fatty acids (SCFAs) in the colonic contents were quantified.

Observations:

· AOM/DSS significantly decreased levels of propionic acid, isobutyric acid, isovaleric acid, and valeric acid;

· After XLJDD treatment, these SCFA levels were significantly elevated;

· Acetic acid and butyric acid did not show significant changes, but a trend of increase was observed with XLJDD;

· Oxaliplatin treatment showed no significant effect on SCFA levels.

Moreover:

· Low-dose XLJDD promoted propionate and isobutyrate production;

· High-dose XLJDD favored isovalerate and valerate accumulation.

XLJDD Modulates Gut Microbiota Dysbiosis in AOM/DSS-Induced Mice：

 

 

Researchers analyzed the effect of XLJDD on gut microbiota diversity and abundance in the AOM/DSS-induced CRC mouse model.

Key findings include:

· XLJDD treatment significantly increased microbial richness and enhanced species diversity;

· PCA and PCoA analyses demonstrated that XLJDD effectively restored microbial community balance disrupted by AOM/DSS;

· Both XLJDD and oxaliplatin treatment led to increased abundance of beneficial bacteria such as Ileibacterium and Lactobacillus, while reducing harmful species like Turicibacter.

These microbial composition shifts are believed to contribute to tumor prevention and immune modulation.

LEfSe Analysis Reveals Key Microbial Biomarkers:

 

 

To identify key microbial taxa and biomarkers across groups, LEfSe (Linear discriminant analysis Effect Size) was applied.

Findings:

· Turicibacter and Dubosiella were highly enriched in the model group and are considered drivers of dysbiosis;

· Under treatment with XLJDD and oxaliplatin, the relative abundance of:

· Lachnospiraceae_NK4A136_group

· Enterorhabdus

· Alistipes

was significantly elevated;

In contrast, harmful taxa such as:

· Turicibacter

· Romboutsia

· Clostridium_sensu_stricto_1

were substantially decreased.These results suggest that XLJDD corrects microbiota imbalances by modulating both the composition and diversity of gut bacteria, which may in turn inhibit CRC development.

XLJDD Alters Metabolites in AOM/DSS-Induced Mice：

 

 

Untargeted metabolomics analysis was conducted on blood plasma and fecal samples.

Key findings are:

· PCA and OPLS-DA analyses revealed significant metabolic differences between the control and model groups;

· Treatment with XLJDD and oxaliplatin effectively reversed 29 differential metabolites disrupted by AOM/DSS;

· KEGG pathway analysis of XLJDD-regulated metabolites showed strong associations with:

· Lipid metabolism

· Amino acid metabolism

· Organic acid metabolism

· Notably, sphingolipid metabolism and glycerophospholipid metabolism were the most strongly linked to XLJDD's therapeutic effects.

This suggests these two pathways may be critical targets of XLJDD in the treatment of CRC.

Correlation Between Gut Microbiota and Clinical Indicators：

 

 

Through co-expression network analysis and Spearman correlation, the study explored the relationships between:

· 29 key metabolites

· 50 gut microbial genera

Key observations are:

· XLJDD’s therapeutic effects may stem from modulating the interaction between specific gut microbes and host metabolites;

· This interaction likely plays a central role in its anti-CRC mechanisms;

· The results offer novel biomarkers and fresh perspectives for further investigation into XLJDD as a colorectal cancer therapy.

Article Summary

1. XLJDD reduced colonic tumor burden and the colon weight/length ratio, improved body weight, colon length, and overall survival in mice;

2. XLJDD lowered inflammatory cytokine levels in serum and downregulated β-catenin, COX-2, and iNOS expression in colorectal tissue, demonstrating anti-inflammatory effects

3. XLJDD improved gut microbiota balance, boosted SCFA production (e.g., butyrate and isovalerate), and enhanced sphingolipid and glycerophospholipid metabolism

4. XLJDD increased beneficial microbes such as Enterorhabdus and Alistipes.

Conclusion

This study combines traditional Chinese medicine theory with modern analytical techniques including:

· Non-targeted TCM component analysis

· 16S microbiome profiling

· Serum and fecal untargeted metabolomics

· SCFA quantification

These integrated methods provide new insights and scientific validation for using TCM in colorectal cancer treatment.For researchers looking to publish high-impact TCM papers, this study offers a powerful, reproducible framework. Just apply it to a different disease model, and you may quickly obtain publishable results.