Nutrients 2015, 7(2), 764-784; doi:10.3390/nu7020764

Intestinal Microbial Dysbiosis and Colonic Epithelial Cell Hyperproliferation by Dietary α-Mangostin is Independent of Mouse Strain

 

Fabiola Gutierrez-Orozco 1,2, Jennifer M. Thomas-Ahner 2,3, Jeffrey D. Galley 2,4, Michael T. Bailey 2,4, Steven K. Clinton 1,2,3,5, Gregory B. Lesinski 1,2,3,5 and Mark L. Failla 1,2,3,*

1Human Nutrition Program, Department of Human Sciences, 2Food Innovation Center, 3Comprehensive Cancer Center, 4Division of Oral Biology, College of Dentistry 5Department of Internal Medicine

The Ohio State University, Columbus, OH 43210, USA

 

Abstract

Beverages and supplements prepared from mangosteen fruit are claimed to support gut health and immunity, despite the absence of supporting evidence from clinical trials. We recently reported that α-mangostin (α-MG), the most abundant xanthone in mangosteen fruit, altered the intestinal microbiome, promoted dysbiosis, and exacerbated chemically-induced colitis in C57BL/6J mice.1 The objective of this subsequent study was to determine whether induction of dysbiosis by dietary α-MG was unique to the C57BL/6J strain or represented a more generic response to chronic intake of the xanthone on the gut microbiota of mice. C3H, Balb/c, Nude FoxN1nu, and C57BL/6J mice, each possessing unique microbiomes, were fed standard diet or diet containing 0.1% α-MG for four weeks. Dietary α-MG significantly altered the cecal and colonic microbiota in all four strains of mice, promoting a reduction in generally assumed beneficial bacterial groups while increasing the abundance of pathogenic bacteria. Consumption of α-MG was associated with reduced abundance of Firmicutes and increased abundance of Proteobacteria. The abundance of Lachnospiraceae, Ruminococcaceae, and Lactobacillaceae was reduced in α-MG-fed mice, while that of Enterobacteriaceae and Enterococcaceae was increased. Dietary α-MG also was associated with increased proliferation of colonic epithelial cells, infiltration of immune cells and increased fluid content in stool. These results suggest that ingestion of pharmacologic doses of xanthones in mangosteen-containing supplements may adversely alter the gut microbiota and should be used with caution.

PMID: 25621505

 

Supplement

Mangosteen is the fruit produced by the Garcinia mangostana tree. Native to Southeast Asia, mangosteen is known there as “the queen of fruits” because of its pleasing organoleptic properties. It also has been used in traditional medicine for the treatment of inflammation, wounds and other ailments. The bioactivities of mangosteen have been associated with a family of polyphenolic compounds known as xanthones, mainly present in the pericarp of the fruit. Among all mangosteen xanthones, α-mangostin (α-MG) is the most abundant and by far the most studied compound in mangosteen fruit. Interest in the bioactivities of this xanthone has significantly increased in recent years as the number of publications describing its anti-cancer, anti-microbial, anti-obesogenic, anti-inflammatory and other health promoting activities continue to increase. In addition, dietary supplements containing mangosteen extracts have become quite popular in the North American and European markets. Such products are usually marketed for their gastrointestinal and immune health benefits, despite extremely limited evidence to support such claims in humans.

Our initial in vitro study showed that α-MG attenuated the inflammatory response in human and rodent cancer cell lines. In contrast, the xanthone stimulated the secretion of pro-inflammatory molecules by non-transformed human derived macrophages were use.2 Dietary α-MG also exacerbated colonic inflammation and injury, promoted greater systemic inflammation and increased recruited numbers of inflammatory cells to the colon when orally administered to C57BL/6J mice in conjunction with a chemically-induced colitis in comparison to colitic mice fed a standard semi-purified diet without α-MG. Several other phytochemicals and plant extracts generally assumed to be health-promoting have been reported to exacerbate DSS-induced colitis in mice. These include dietary luteolin, tomato lycopene, green tea polyphenolic extracts and flaxseed.3-6 We also found dysbiosis in the colon and cecum in non-colitic mice fed diet containing the xanthone.1 These results were unexpected as we had found that the same dose of dietary α-mangostin attenuated the growth of tumors resulting from sub-cutaneous injection of HT-29 human colonic carcinoma cells in nude mice.7

Because host genotype is one of many factors affecting the host inflammatory response as well as the composition of the gut microbiota, the primary objective of the more recent study was to determine whether the induction of dysbiosis by dietary α-MG in mice that were not administered the colonic pro-inflammatory insult was dependent on mouse genotype or represented a generic response of different strains to the xanthone. Otherwise healthy female C3H, Balb/c, Nude FoxN1nu, and C57BL/6J mice were fed semi-purified diet containing 0.1% α-MG for 4 weeks. Tissue-associated bacteria in the cecum and colon, as well as the histological profiles of the colon, of these mice were analyzed. Our results clearly show that feeding a diet containing 0.1% α-MG for 4 caused increased fluid content in stools in the absence of alterations in food intake or body weight. Moreover, chronic consumption of the diet with α-MG induced dysbiosis in the colon and cecum of all four murine strains (Figure 1). Greater colonic epithelial cell proliferation and infiltration of T lymphocytes and macrophages were also observed in the colon of mice fed diet containing α-MG.

 

FIG1

Figure 1. Changes in the bacterial profile (expressed as relative abundance) induced by α-mangostin at the phylum level in the colon of C57BL/6J, Balb/c, C3H and FoxN1nu mice.

 

Our results suggest that ingestion of α-MG may adversely impact the gut microbiota. These alterations resemble some of the changes present in the microbiota of individuals with IBD.8-11 The amount of α-MG tested in our pre-clinical study are relevant to that which can be ingested by humans commercially available mangosteen-containing beverages and supplements. Thus, we have suggested that consumption of pharmacological doses of xanthones in mangosteen-containing supplements may have unintended consequences on the gut microbiota and their use should be considered with caution pending further information.1,2 Such caution is supported by the observation that chronic ingestion of juices containing mangosteen fruit was associated with increases in several pro-inflammatory markers in the serum of human subjects in the only two studies reported to date.12,13

There are several limitations of our studies that provide a framework for further investigations. The adverse impact of α-MG on the microbiota and the large intestinal epithelium was investigated using a single model of colitis. Whether α-MG exacerbates colonic inflammation and injury in other models of colitis merits consideration. In addition, only a single dose of α-MG was used. Mangosteen pericarp contains more than 50 distinct xanthones. The relative bioactivities of these xanthones alone or in combination remain to be determined. Whether the different xanthones interact in additive, synergistic, or antagonistic manners is unknown. It also is possible that the bioactivities of mangosteen xanthones are mediated by inducing an adaptive stress or “hormetic” response.14,15 Therefore, higher concentrations of these poorly absorbed compounds or their bioactive metabolites within the gastrointestinal tract may not necessarily be beneficial. Future investigations should evaluate the effects of lower doses of α-MG, other xanthones, and mixtures of mangosteen xanthones on the gut microbiome and colonic epithelial function in healthy animals, as well as those with gut pathologies.  The actual amount of xanthones in the gut lumen that are available for uptake and metabolism by intestinal epithelial cells and the microbiota may be dependent on the type of ingested product, namely, milled pericarp, xanthone-rich extracts, or pure xanthones. Xanthone release from pericarp during digestion has not been addressed. Finally, because fiber also is present in beverages and other products containing milled pericarp, the extent to which this fiber is fermented in the large intestine may modify the amounts and metabolism of xanthones within the colonic lumen.

 

References

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Acknowledgements: This study was supported by grants from the Molecular Carcinogenesis and Chemoprevention Program and the Food Innovation Center at The Ohio State University (Gregory B. Lesinski, Steven K. Clinton, and Mark L. Failla), the Ohio Agriculture Research and Development Center (Fabiola Gutierrez-Orozco and Mark L. Failla) and the Conacyt Doctoral Scholarship of Mexico and OSU College of Education and Human Ecology Dissertation Fellowship (Fabiola Gutierrez-Orozco).

 

Contact: Mark Failla – failla.3@osu.edu; 614-560-9401

 

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