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Updated: Feb 16, 2026
Fecal micro RNA Isolation
Published on: October 28, 2020
Faecal microRNAs: indicators of imbalance at the host-microbe interface?
G M Moloney1, M F Viola1, A E Hoban1,2
11 Department of Anatomy and Neuroscience, University College Cork, Western Gateway Building, Western Rd., Cork, Ireland.
Abstract:
The enteric microbiota is characterised by a balance and composition that is unique to the host. It is important to understand the mechanisms through which the host can maintain the composition of the gut microbiota. MicroRNAs (miRNA) are implicated in intercellular communication and have been isolated from bodily fluids including stool. Recent findings suggest that miRNA produced by the host's intestinal epithelial cells (IECs) participate in shaping the microbiota. To investigate whether miRNA expression was influenced by the gut microbiota we measured the expression of miRNAs expressed by intestinal epithelial cells in faeces. Specifically, we measured miRNA expression in faeces from germ-free (GF) and conventional mice and similarly in a rat model of antibiotic-mediated depletion of the gut microbiota control rats. In adult male GF and conventional mice and adult Sprague Dawley (SD) rats were treated with a combination of antibiotics for 8 weeks; total RNA was extracted from faecal pellets taken at week 0, 2, 4, 6 week 8 and the expression of let-7b-3p, miR-141-3p, miR-200a-3p and miR-1224-5p (miRNAs known to be expressed in IECs) were measured relative to U6 at each time point using qRT-PCR. In GF animals the expression of let-7b, miR-141 and miR-200a in faeces was lower compared to conventional mice. Following antibiotic-mediated depletion of gut microbiota, rats showed two divergent profiles of miRNA expression. Following two weeks of antibiotic treatment, the expression of let-7b and miR-1224 dropped significantly and remained low for the remainder of the study. The expression of miR-200a and miR-141 was significantly higher at week 2 than before antibiotic treatment commenced. Subsequently, the expression of miR-200a and miR-141 decreased at week 4 and continued to decrease at week 6. This data demonstrates that miRNAs can be used as an independent, non-invasive marker of microbial fluctuations along with gut pathology in the intestine.
Insights
Host intestinal epithelial cell microRNAs (miRNAs) are influenced by gut microbiota composition. Fecal miRNA levels can serve as non-invasive biomarkers for tracking gut microbial changes and associated pathologies.
Area of Science:
- Gastroenterology
- Molecular Biology
- Microbiome Research
Background:
- The gut microbiota's composition is host-specific and crucial for intestinal health.
- MicroRNAs (miRNAs) are key regulators of gene expression and are found in stool, suggesting a role in host-microbe communication.
- Intestinal epithelial cell (IEC)-derived miRNAs may influence gut microbiota homeostasis.
Purpose of the Study:
- To investigate the influence of gut microbiota on the expression of specific miRNAs in the intestine.
- To determine if fecal miRNA expression can serve as a non-invasive marker for microbial fluctuations and gut health.
Main Methods:
- Measured fecal expression of IEC-specific miRNAs (let-7b-3p, miR-141-3p, miR-200a-3p, miR-1224-5p) using qRT-PCR in germ-free and conventional mice.
- Analyzed miRNA expression in rats undergoing antibiotic-induced gut microbiota depletion over 8 weeks.
Main Results:
- Germ-free mice exhibited lower fecal levels of let-7b, miR-141, and miR-200a compared to conventional mice.
- Antibiotic treatment in rats led to distinct miRNA expression profiles: let-7b and miR-1224 decreased, while miR-200a and miR-141 initially increased then decreased.
- Fecal miRNA levels correlated with microbial status and potential gut pathology.
Conclusions:
- Gut microbiota composition significantly impacts the expression of intestinal epithelial cell-derived miRNAs.
- Fecal miRNAs represent a promising non-invasive biomarker for monitoring gut microbiota dynamics and intestinal health.
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