One Month of Classic Therapeutic Ketogenic Diet Decreases Short Chain Fatty Acids Production in Epileptic Patients

Cinzia Ferraris, Erika Meroni, Maria Cristina Casiraghi, Anna Tagliabue, Valentina De Giorgis, Daniela Erba, Cinzia Ferraris, Erika Meroni, Maria Cristina Casiraghi, Anna Tagliabue, Valentina De Giorgis, Daniela Erba

Abstract

Ketogenic diet (KD), a high fat and very low carbohydrates diet, is used worldwide for the treatment of drug resistant epilepsy but, due to its composition, it might exert an impact on gut health. Even though data of KD effects on intestinal microbiota changes are recently emerging, its influence on the gut environment has been scarcely addressed so far. The aim of this study was to investigate whether 1 month of KD affects the gut environment in epileptic patients, by analyzing short chain fatty acids (SCFA) production and fecal water toxicity. A total of seven patients were enrolled. Stool samples were collected before (T0) and after 1 month of KD (4:1 ketogenic ratio) (T1). SCFA were determined by GC-FID and fecal water toxicity in Caco-2 cell culture by comet assay. Concentrations of SCFA significantly decreased after KD (p < 0.05): in particular, we found a 55% reduction of total SCFA level, a 64% reduction of acetate, 33% of propionate, and 20% of butyrate (p < 0.05). Cytotoxicity of fecal water extracted from stool samples was not significantly altered by diet, while genotoxicity was slightly decreased after KD (p < 0.05). Genotoxicity values were consistent with data previously obtained from a healthy Italian population. The present study suggests that 1 month of KD significantly reduce SCFA production. Since SCFA produced by gut microbiota exert many health promoting effects on either the gut environment or human metabolism, these results open a new branch of investigation into KD effects.

Keywords: epilepsy; fecal water toxicity; gut environment; ketogenic diet; short chain fatty acids.

Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Copyright © 2021 Ferraris, Meroni, Casiraghi, Tagliabue, De Giorgis and Erba.

References

    1. Martin-McGill KJ, Jackson CF, Bresnahan R, Levy RG, Cooper PN. Ketogenic diets for drug-resistant epilepsy. Cochrane Database Syst Rev. (2018) 11:CD001903. 10.1002/14651858.CD001903.pub4
    1. Kossoff EH, Zupec-Kania BA, Auvin S, Ballaban-Gil KR, Christina Berqqvist AG, Blackford R, et al. . Optimal clinical management of children receiving dietary therapies for epilepsy:Updated recommendations of the International Ketogenic Diet Study Group. Epilepsia Open. (2018) 3:175–92. 10.1002/epi4.12225
    1. Newell C, Bomhof MR, Reimer RA, Hittel DS, Rho JM, Shearer J. Ketogenic diet modifies the gut microbiota in a murine model of autism spectrum disorder. Mol Autism. (2016) 7:37. 10.1186/s13229-016-0099-3
    1. Ma D, Wang AC, Parikh I, Green SJ, Hoffman JD, Chlipala G, et al. . Ketogenic diet enhances neurovascular function with altered gut microbiome in young healthy mice. Sci Rep. (2018) 8:6670. 10.1038/s41598-018-25190-5
    1. Olson CA, Vuong HE, Yano JM, Liang QY, Nusbaum DJ, Hsiao EY. The gut microbiota mediates the anti-seizure effects of the ketogenic diet. Cell. (2018) 173:1728–41. 10.1016/j.cell.2018.04.027
    1. Lindefeldt M, Eng A, Darban H, Bjerkner A, Zetterstrom CK, Allander T, et al. . The ketogenic diet influences taxonomic and functional composition of the gut microbiota in children with severe epilepsy. NPJ Biofilms Microbiomes. (2019) 5:5. 10.1038/s41522-018-0073-2
    1. Swidsinski A, Dörffel Y, Loening-Baucke V, Gille C, Göktas O, Reißhauer A, et al. . Reduced mass and diversity of the colonic microbiome in patients with multiple sclerosis and their improvement with ketogenic diet. Front Microbiol. (2017) 8:1141. 10.3389/fmicb.2017.01141
    1. Tagliabue A, Ferraris C, Uggeri F, Trentani C, Bertoli S, de Giorgis V, et al. . Short-term impact of a classical ketogenic diet on gut microbiota in GLUT1 Deficiency Syndrome: a 3-month prospective observational study. Clin Nutr ESPEN. (2017) 17:33–7. 10.1016/j.clnesp.2016.11.003
    1. Xie G, Zhou Q, Qiu C, Dai WK, Wang HP, Li YH, et al. . Ketogenic diet poses a significant effect on imbalanced gut microbiota in infants with refractory epilepsy. World J Gastroenterol. (2017) 23:6164–71. 10.3748/wjg.v23.i33.6164
    1. Zhang Y, Zhou S, Zhou Y, Yu L, Zhang L, Wang Y. Altered gut microbioma composition in children with refractory epilepsy after ketogenic diet. Epilepsy Res. (2018) 145:163–8. 10.1016/j.eplepsyres.2018.06.015
    1. Caprio M, Infante M, Moriconi E, Armani A, Fabbri A, Mantovani G, et al. . Very-low calorie ketogenic diet (VLCKD) in the management of metabolic diseases: systematic review and consensus statement from the Italian Society of Endocrinology (SIE). J Endocrinol Inv. (2019) 42:1365–86. 10.1007/s40618-019-01061-2
    1. Huazano-Garcia A, Lopez MG. Metabolism of short chain fatty acids in the colon and faeces of mice after a supplementation of diets with agave fructans. In: Valenzuela-Baez R. editor. Lipid Metabolism. Rijeka, Croatia: InTech; (2013). p. 163. 10.5772/51248
    1. Wang HB, Wang PY, Wang X, Wan YL, Liu YC. Butyrate enhances intestinal epithelial barrier function via up-regulation of tight junction protein Claudin-1 transcription. Dig Dis Sci. (2012) 57:3126–35. 10.1007/s10620-012-2259-4
    1. Wu X, Wu Y, He L, Wu K, Wang X, Liu Z. Effect of the intestinal microbial metabolite butyrate on the development of colorectal cancer. J Cancer. (2018) 9:2510–7. 10.7150/jca.25324
    1. Segain J-P, de la Blétière DR, Bourreille A, Leray V, Gervois N, Rosales C, et al. . Butyrate inhibits inflammatory responses through NFkappaB inhibition:implications for Crohn's disease. Gut. (2000) 47:397–409. 10.1136/gut.47.3.397
    1. Morrison DJ, Preston MT. Formation of short chain fatty acids by the gut microbiota and their impact on human metabolism. Gut Microbes. (2016) 7:189–200. 10.1080/19490976.2015.1134082
    1. Venturi M, Hambly RJ, Glinghammer B, Rafter JJ, Rowland IR. Genotoxicity activity in human fecal water and the role of bile acids:a study using the alkaline comet assay. Carcinogenesis. (1997) 18:2353–9. 10.1093/carcin/18.12.2353
    1. Cross AJ, Greetham HL, Pollack JRA, Rowland IR, Bingham S.A. Variability in fecal water genotoxicity, determined using the Comet assay, is independent of endogenous N-nitroso compound formation attributed to red meat consumption. Environ Mol Mutagen. (2006) 47:179–84. 10.1002/em.20181
    1. Erba D, Soldi S, Malavolti M, Aragone G, Meynier A, Vinoy S, et al. . Fecal water genotoxicity in healthy free-living young Italian people. Food Chem Toxicol. (2014) 64:104–9. 10.1016/j.fct.2013.11.019
    1. Woods JA, Dunne C, Collins JK, Shanahan F, O'Brien NM. Genotoxicity of fecal water in a free-living Irish population. Nutr Cancer. (2002) 42:62–9. 10.1207/S15327914NC421_9
    1. Bertoli S, Giulini Neri I, Trentani C, Ferraris C, De Amicis R, Battezzati A, et al. . Short-term effects of ketogenic diet on anthropometric parameters, body fat distribution, and inflammatory cytokine production in GLUT1 deficiency syndrome. Nutrition. (2015) 31:981–7. 10.1016/j.nut.2015.02.017
    1. AOAC . Official methods of analysis Proximate Analysis and Calculations Moisture (M) - item 105. Association of Analytical Communities, Gaithersburg, MD, 17th edition, (2006) Reference data: Method 934.01.
    1. Weaver GA, Tangel CT, Krause JA, Parfitt MM, Jenkins PL, Rader JM, et al. . Acarbose enhances human colonic butyrate production. J Nutr. (1997) 127:717–23. 10.1093/jn/127.5.717
    1. Borgo F, Riva A, Benetti A, Casiraghi MC, Bertelli S, Garbossa S, et al. . Microbiota in anorexia nervosa:the triangle between bacterial species, metabolites and psychological tests. PLoS ONE. (2017) 12:e0179739. 10.1371/journal.pone.0179739
    1. Sambuy Y, de Angelis I, Ranaldi G, Scarino ML, Stammati A, Zucco F. The Caco-2 cell line as a model of the intestinal barrier:influence of cell and culturerelated factors on Caco-2 cell functional characteristics. Cell Biol Toxicol. (2005) 21:1–26. 10.1007/s10565-005-0085-6
    1. Meroni E, Papini N, Criscuoli F, Casiraghi MC, Massaccesi L, Basilico N, et al. . Metabolic responses in endothelial cells following exposure to ketone bodies. Nutrients. (2018) 10:250. 10.3390/nu10020250
    1. Petersen C, Round J. Defining dysbiosis and its influence on host immunity and disease. Cell Microbiol. (2014) 16:1024–33. 10.1111/cmi.12308
    1. Borghi E, Borgo F, Severgnini M, Savini MN, Casiraghi MC, Vignoli A. Rett syndrome: a focus on gut microbiota. Int J Mol Sci. (2017) 18:344. 10.3390/ijms18020344
    1. Yamada T, Shimizu K, Ogura H, Asahara T, Nomoto K, Yamakawa K, et al. . Rapid and sustained long-term decrease of fecal short-chain fatty acids in critically ill patients with systemic inflammatory response syndrome. JPEN J Parenter Enteral Nutr. (2015) 39:569–77. 10.1177/0148607114529596
    1. Chassard C, Lacroix C. Carbohydrates and the human gut microbiota. Curr Opin Clin Nutr Metab Care. (2013) 16:453–60. 10.1097/MCO.0b013e3283619e63
    1. Kleesen B, Hartmann L, Blaut M. Oligofructose and long chain inulin:influence on the gut microbial ecology of rats associated with a human fecal flora. Br J Nutr. (2001) 86:375–82. 10.1079/BJN2001403
    1. Koropatkin NM, Cameron EA, Martens EC. How glycan metabolism shapes the human gut microbiota. Nat Rev Microbiol. (2012) 10:323–35. 10.1038/nrmicro2746
    1. de Kok TMCM, van Maanen JMS. Evaluation of fecal mutagenicity and colon cancer risk. Mutat Res. (2000) 463:53–101. 10.1016/S1383-5742(00)00003-X
    1. Mai V, McCrary QM, Sinha R, Glei M. Associations between dietary habits and body mass index with gut microbiota composition and fecal water genotoxicity:an observational study in African American and Caucasian American volunteers. Nutr J. (2009) 8:49. 10.1186/1475-2891-8-49
    1. Reddy BS, Shamsudden AKM. Nutritional factors and colon cancer. Crit. Rev Food Sci. Nutr. (1995) 35:175–190. 10.1080/10408399509527698
    1. Hori T, Matsumoto K, Sakaitani Y, Sato M, Morotomi M. Effect of dietary deoxycholic acid and cholesterol on fecal steroid concentration and its impact on the colonic crypt cell proliferation in azoxymethane-treated rats. Cancer Lett. (1998) 124:79–84. 10.1016/S0304-3835(97)00452-7
    1. Imray CH, Radley S, Davis A, Barker G, Hendrickse C, Donovan IA, et al. . Faecal unconjugated bile acids in patients with colorectal cancer or polyps. Gut. (1992) 33:1239–45. 10.1136/gut.33.9.1239
    1. Stadler J, Stern HS, Sing-Yeung KA, McGuire V, Furrer R, Marcon N, et al. . Effect of high fat consumption on cell proliferation activity of colorectal mucosa and on soluble faecal bile acids. Gut. (1988) 29:1326–31. 10.1136/gut.29.10.1326
    1. Powolny A, Xu J, Loo G. Deoxycholate induces DNA damage and apoptosis in human colon epithelial cells expressing either mutant or wild-type p53. Int J Biochem Cell Biol. (2001) 33:193–203. 10.1016/S1357-2725(00)00080-7
    1. Osswald K, Becker TW, Grimm M, Jahreis G, Pool-Zobel BL. Inter- and intra-individual variation of fecal water genotoxicity in human colon cells. Mutat Res. (2001) 472:59–70. 10.1016/S1383-5718(00)00113-3
    1. Erba D, Casiraghi MC, Martinez-Conesa C, Goi G, Massaccesi L. Isoflavone supplementation reduces DNA oxidative damage and increases O-β-N-acetyl-D-glucosaminidase activity in healthy women. Nutr Res. (2012) 32:233–40. 10.1016/j.nutres.2012.03.007
    1. Ferraris C, Elli M, Tagliabue A. Gut microbiota for health:how can diet maintain a healthy gut microbiota? Nutrients. (2020) 12:3596. 10.3390/nu12113596
    1. Meroni E. Metabolic effects of dietary approaches: ketone bodies and ketogenic diet [Doctoral Thesis]. Milan, IT: University of Milan; (2018).

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