Therapeutic Evaluation of Fecal Microbiota Transplantation in an Interleukin 10-Deficient Mouse Model

Podsumowanie

The interaction of genetic susceptibility, mucosal immunity, and intestinal microecological environment is involved in the pathogenesis of inflammatory bowel disease (IBD). In this study, we applied fecal microbiota transplantation to IL-10 deficient mice and investigated its impact on colonic inflammation and heart function.

Streszczenie

With the development of microecology in recent years, the relationship between intestinal bacteria and inflammatory bowel disease (IBD) has attracted considerable attention. Accumulating evidence suggests that dysbiotic microbiota plays an active role in triggering or worsening the inflammatory process in IBD and that fecal microbiota transplantation (FMT) is an attractive therapeutic strategy since transferring a healthy microbiota to IBD patient could restore the appropriate host-microbiota communication. However, the molecular mechanisms are unclear, and the efficacy of FMT has not been very well established. Thus, further studies in animal models of IBD are necessary. In this method, we applied FMT from wild-type C57BL/6J mice to IL-10 deficient mice, a widely used mouse model of colitis. The study elaborates on collecting fecal pellets from the donor mice, making the fecal solution/suspension, administering the fecal solution, and monitoring the disease. We found that FMT significantly mitigated the cardiac impairment in IL-10 knockout mice, underlining its therapeutic potential for IBD management.

Wprowadzenie

The human intestinal micro-ecosystem is extremely complex, with more than 1000 species of bacteria in the intestine of a healthy person¹. The intestinal flora is involved in maintaining the normal physiological functions of the intestine and the immune response and has an inseparable relationship with the human body. Accumulating evidence suggests that the intestinal microbiome constitutes the last human organ, which is part of the human body, not just a group of parasites². A 'healthy' symbiotic relationship between the gut microbiota, their metabolites, and the host immune system established in early life is cr....

Protokół

All procedures performed on animals were approved by the Institutional Animal Care and Use Committee of the University of Texas Medical Branch at Galveston (Protocol # 1512071A).

1. Collection of fresh fecal pellets

1. Prepare sterile paper towels, blunt-end forceps, and 50 mL conical tubes.
   1. Place some paper towels and forceps in separate autoclave bags and autoclave them at 180 °C in dry heat for 30 min. Use sterile conical tubes as well. Weigh the conical tubes and write down their weight on the tubes.
2. Turn on the biosafety cabinet in the animal room.
3. Take an aut....

Wyniki

We performed healthy donor FMT 3 times (once a month for 3 months) on 2-month old C57BL/6J wild type (WT) and IL-10 knockout mice. Age-matched C57BL/6J mice (age difference should be <2 months) served as the fecal donors and fresh fecal pellets were used each time. EIA assays revealed that BNP was markedly elevated in the plasma of IL-10-deficiency mice and that healthy donor FMT significantly mitigated the increase in BNP levels (Figure 1A, n = 5, p < 0.05). Echocardiograph.......

Dyskusje

As an innovative investigational treatment, FMT has become a hot topic in the treatment of various disorders in recent years since dysbiosis of the commensal microbiota is implicated in the pathogenesis of multiple human diseases, including IBD, obesity, diabetes mellitus, autism, heart disease, and cancer²⁶. Although the mechanism has not been determined, it is believed that FMT works by building a new biological flora and preventing the loss of residual bacteria. The method presented herein adop.......

Materiały

Name	Company	Catalog Number	Comments
BD Syringe, 1 mL	Fisher Scientific	14-829-10F
Blunt end forceps	Knipex	926443
Brain natriuretic peptide EIA kit	Sigma	RAB0386
C57BL/6J mice	Jackson Lab	000664
Centrifuge	Eppendorf	5415R
Conical tubes	ThermoFisher	339650
Curved feeding Needles	Kent Scientific	FNC-20-1.5-2
GLH-115 homogenizer	Omni International	GLH-115
Glycerol	MilliporeSigma	G5516
IL-10 knockout mice	Jackson Lab	004366
Isoflurane	Piramal Critical care	NDC66794-017-10
USP normal saline	Grainger	6280
Vaporizer	Euthanex Corp.	EZ-108SA