Borowicki, A.

Brenner-Weiss, G.

Glei, M.

Gusko, M.

Pool-Zobel, B.L.

Scharlau, D.

Scheu, K.

Stein, K.

Pool-Zobel, B. L.

KITopen

fs0 5 10 15 20050100150**fs blankfs US-aleuronefs EU-aleurone **concentration [%]cellnumber[%]SCFA0 5 10 15 20050100150fs blankfs US-aleuronefs EU-aleuroneconcentration [%]cellnumber[%]butyrate0 5 10 15 20050100150fs blankfs US-aleuronefs EU-aleuroneconcentration [%]cellnumber[%]********** ******0 5 10 15 20050100150*fs blankfs US-aleuronefs EU-aleuroneconcentration [%]cellnumber[%]***0 5 10 15 20050100150***fs blankfs US-aleuronefs EU-aleuroneconcentration [%]cellnumber[%]*****0 5 10 15 20050100150***fs blankfs US-aleuronefs EU-aleuroneconcentration [%]cellnumber[%]**************** ***0 5 10 15 20050100150*******fs blankfs US-aleuronefs EU-aleuroneconcentration [%]cellnumber[%]**0 5 10 15 20050100150*** ****** *****fs blankfs US-aleuronefs EU-aleurone**fs blank fs EU-aleurone fs US-aleuroneconcentration [%]cellnumber[%]***0 5 10 15 20050100150** *******fs blankfs US-aleuronefs EU-aleuroneconcentration [%]cellnumber[%]*24 h48 h72 h****fs0 5 10 15 20050100150fs branfs flourfs aleuronefs blank*** ****concentration [%]cellnumber[%]SCFA0 5 10 15 20050100150fs flourfs aleuronefs blankconcentration [%]cellnumber[%]butyrate0 5 10 15 20050100150fs flourfs aleuronefs blankconcentration [%]cellnumber[%]**********************0 5 10 15 20050100150*fs branfs flourfs aleuronefs blankconcentration [%]cellnumber[%]0 5 10 15 20050100150*** ****** ****fs flourfs aleuronefs blankconcentration [%]cellnumber[%]**0 5 10 15 20050100150*********fs flourfs aleuronefs blankconcentration [%]cellnumber[%]**************0 5 10 15 20050100150*****************fs branfs flourfs aleuronefs blankconcentration [%]cellnumber[%]***0 5 10 15 20050100150*** ****** *******fs flourfs aleuronefs blankfs blank fs aleurone fs flour fs branconcentration [%]cellnumber[%]0 5 10 15 20050100150*** ****** **********fs flourfs aleuronefs blankconcentration [%]cellnumber[%]24 h48 h72 hAnke Borowicki ,Katrin Stein , Daniel Scharlau ,Michael Glei , Kerstin Scheu ,Gerald Brenner-Weiss ,Michael Gusko , Beatrice L. Pool-Zobel1 1 1 1 2 23 1123Department of Nutritional Toxicology, Institute for Nutrition, Friedrich-Schiller-University Jena, Dornburger Str. 24, 07743 Jena, GermanyResearch Centre Karlsruhe, Institute for Technical Chemistry, Group for Water- and Geotechnology, Hermann-von-Helmholtz-Platz 1, 76344 Eggenstein-Leopoldshafen, GermanyKampffmeyer Food Innovation GmbH, Trettaustr. 32-34, 21107 Hamburg, GermanyGut flora-mediated fermentation of wheat aleurone independently from variety results in reduced level of tumour promoting DCAand higher levels of SCFA especially butyrate, which inhibits growth of human adenocarcinoma cells. The fact that complex fs weremore cytotoxic than corresponding synthetic mixtures of SCFA and butyrate point to the involvement of additional growth inhibitoryeffects of other bacterial metabolites. Most of the activities of whole meal and of bran are due to their contents of aleurone.Wheat aleurone, whole meal wheat flour and wheat bran were digested and fermented . Fermentation supernatants (fs) wereanalysed for SCFA DCA, ammonia and H O . Corresponding mixtures of SCFA and theindividual substancebutyrate were prepared.HT29 adenocarcinoma cells were treated for 24-72 h with butyrate, mixtures or complex fs. Cell survival was determined byquantifying fluorescence of DAPI-labelled DNA.in vitro, 2 2ResultsDCA [µM] H2O2 [µM] ammonia [mM]fs blank 35.4 87.4 10.9fs aleurone 10.3 90.0 18.5fs flour 2.6 74.7 11.6fs bran 9.1 83.1 14.6DCA [µM] H2O2 [µM] ammonia [mM]fs blank 46.65 77.6 11.9fs EU-aleurone 5.04 100.6 16.6fs US-aleurone 5.93 106.5 21.3Acknowedgements For funding we would like to thank the Federal Ministry of Education and Research, Germany (BMBF 0313829A).Figure 1: (A) Longitudinal section of a wheat grain and focus on aleurone layer.(B) F ifferent wheat samples which were fermented .lours of d in vitro(A) (B)Figure 2: Concentrations and molar ratios of SCFA in complex fs of wheat samples and control(blank) analysed by GC-MS.Table 1: Concentrations of DCA, H O and ammonia in complex fs of wheat samplesand control (blank) analysed by HPLC-MS/MS, FOX2 assay and Berthelot reaction.2 2Figure 3: Concentrations and molar ratios of SCFA in complex fs of EU- and US-aleuroneand control (blank) analysed by GC-MS.Table 2: Concentrations of DCA, H O and ammonia in complex fsand control (blank) analysed by HPLC-MS/MS, FOX2 assay and Berthelot reaction.2 2 of EU- and US-aleuroneFigure 4: Inhibition of cell growth after incubation of HT29 cells with complex fs and corresponding synthetic mixtures of (A) different wheat samples or (B) two wheat aleurone varieties (EU: european, US: american) and control (blank). Shown are means ± SD (n=3) and significantdifferences of respective concentrations of fs or synthetic substances to medium control (p<0.05: *; p<0.01: **; p<0.001: ***).whole meal bran aleuroneBackgroundWheat grain and particularly the aleurone layer (Fig. 1) contain high amountsof dietary fibre. Fermentation of dietary fibre by human gut flora may enhancelevel of short-chain fatty acids (SCFA) which are potentially chemoprotectivee.g. by suppressing the growth of tumour cells. Furthermore lower amountsof potentially tumour promoting products such as deoxycholic acid (DCA),ammonia and hydrogen peroxide (H O ) may be produced.2 2 .GUT FERMENTATION PRODUCTS OF WHEAT ALEURONE SUPPRESSCELL GROWTH AND SURVIVAL OF HUMAN ADENOCARCINOMA CELLSMethodsConclusions(A) (B)fs blank fs aleurone fs flour fs bran020406080100acetate propionate butyratemolarratio[%]fs blank fs aleurone fs flour fs bran020406080100concentration[mM]59-18-23 43-19-3853-11-36 46-17-3726835064Fermented wheat samples suppressed growth of tumour cells. Fs inhibited cell growth more than synthetic mixtures.Growth inhibitory activity of SCFA was mainly caused by butyrate (Fig. 4).fs blank fs EU-aleurone fs US-aleurone020406080100concentration[mM]fs blank fs EU-aleurone fs US-aleurone020406080100acetate propionate butyratemolarratio[%]54-23-23 43-15-42 43-16-41204859Fermented wheat samples contained2-3 fold higher amounts ofSCFA than control (Fig. 2, 3).Fermented wheat samples containedreduced levels of DCA. H O productionwas not influenced whereas moreammonia was produced (Tab. 1, 2).2 2

Gut fermentation products of wheat aleurone suppress cell growth and survival of human adenocarcinoma cells

https://publikationen.bibliothek.kit.edu/220071312/3815194

Gut fermentation products of wheat aleurone suppress cell growth and survival of human adenocarcinoma cells

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