Gut microbiota composition in male rat models under different nutritional status and physical activity and its association with serum leptin and ghrelin levels

María Isabel Queipo-Ortuño, Luisa María Seoane, Mora Murri, María Pardo, Juan Miguel Gomez-Zumaquero, Fernando Cardona, Felipe Casanueva, Francisco J Tinahones, María Isabel Queipo-Ortuño, Luisa María Seoane, Mora Murri, María Pardo, Juan Miguel Gomez-Zumaquero, Fernando Cardona, Felipe Casanueva, Francisco J Tinahones

Abstract

Background: Several evidences indicate that gut microbiota is involved in the control of host energy metabolism.

Objective: To evaluate the differences in the composition of gut microbiota in rat models under different nutritional status and physical activity and to identify their associations with serum leptin and ghrelin levels.

Methods: In a case control study, forty male rats were randomly assigned to one of these four experimental groups: ABA group with food restriction and free access to exercise; control ABA group with food restriction and no access to exercise; exercise group with free access to exercise and feed ad libitum and ad libitum group without access to exercise and feed ad libitum. The fecal bacteria composition was investigated by PCR-denaturing gradient gel electrophoresis and real-time qPCR.

Results: In restricted eaters, we have found a significant increase in the number of Proteobacteria, Bacteroides, Clostridium, Enterococcus, Prevotella and M. smithii and a significant decrease in the quantities of Actinobacteria, Firmicutes, Bacteroidetes, B. coccoides-E. rectale group, Lactobacillus and Bifidobacterium with respect to unrestricted eaters. Moreover, a significant increase in the number of Lactobacillus, Bifidobacterium and B. coccoides-E. rectale group was observed in exercise group with respect to the rest of groups. We also found a significant positive correlation between the quantity of Bifidobacterium and Lactobacillus and serum leptin levels, and a significant and negative correlation among the number of Clostridium, Bacteroides and Prevotella and serum leptin levels in all experimental groups. Furthermore, serum ghrelin levels were negatively correlated with the quantity of Bifidobacterium, Lactobacillus and B. coccoides-Eubacterium rectale group and positively correlated with the number of Bacteroides and Prevotella.

Conclusions: Nutritional status and physical activity alter gut microbiota composition affecting the diversity and similarity. This study highlights the associations between gut microbiota and appetite-regulating hormones that may be important in terms of satiety and host metabolism.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1. Representation of the relative daily…
Figure 1. Representation of the relative daily body weight variation (A) and daily food intake (B) in the all study groups (N =  10 rats per group).
The figure (C) shows a 23 h registered activity in ABA and exercise groups.
Figure 2. Serum leptin (A) and grhelin…
Figure 2. Serum leptin (A) and grhelin (B) levels (C) at day 6 of study in all rat groups (N =  10 rats per group).
The Mann-Whitney U test was used to compare a rat group to each other. Different superscript letters are significantly different P<0.05 (Bonferroni Post-hoc test).
Figure 3. Dendrograms of electrophoretic band patterns…
Figure 3. Dendrograms of electrophoretic band patterns obtained in the denaturing gel gradient electrophoresis experiment with universal primers in the fecal samples collected from ABA, control ABA (cABA), exercise and ad libitum rats.
A: Cluster analysis; B: DGGE profiles of fecal samples.

References

    1. Yager J, Anderson AE (2005) Anorexia nervosa. N Engl J Med 353(14): 1481–1488.
    1. Affenito SG, Dohm FA, Crawford PB, Daniels SR, Striegel-Moore RH (2002) Macronutrient intake in anorexia nervosa: The National Heart, Lung, and Blood Institute Growth and Health Study. J Pediatr 141(5): 701–705.
    1. Payne AN, Chassard C, Banz Y, Lacroix C (2012) The composition and metabolic activity of child gut microbiota demonstrate differential adaptation to varied nutrient loads in an in vitro model of colonic fermentation. FEMS Microbiol Ecol 80(3): 608–623.
    1. Kowalska I, Karczewska-Kupczewska M, Strączkowski M (2011) Adipocytokines, gut hormones and growth factors in anorexia nervosa. Clin Chim Acta 412(19–20): 1702–1711.
    1. Hebebrand J, Muller TD, Holtkamp K, Herpertz-Dahlmann B (2007) The role of leptin in anorexia nervosa: clinical implications. Mol Psychiatry 12(1): 23–35.
    1. Boakes RA (2007) Self-starvation in the rat: running versus eating. Span J Psychol 10(2): 251–257.
    1. Bulik CM, Berkman ND, Brownley KA, Sedway JA, Lohr KN (2007) Anorexia nervosa treatment: a systematic review of randomized controlled trials. Int J Eat Disord 40(4): 310–320.
    1. Nedvídková J, Krykorková I, Barták V, Papezová H, Gold PW, et al. (2003) Loss of meal-induced decrease in plasma ghrelin levels in patients with anorexia nervosa. J Clin Endocrinol Metab 88(4): 1678–1682.
    1. Castaneda TR, Tong J, Datta R, Culler M, Tschöp MH (2010) Ghrelin in the regulation of body weight and metabolism. Front Neuroendocrinol 31(1): 44–60.
    1. De Vriese C, Delporte C (2008) Ghrelin: a new peptide regulating growth hormone release and food intake. Int J Biochem Cell Biol 40(8): 1420–1424.
    1. Broglio F, Prodam F, Riganti F, Muccioli G, Ghigo E (2006) Ghrelin: from somatotrope secretion to new perspectives in the regulation of peripheral metabolic functions. Front Horm Res 35: 102–114.
    1. Ducrotte P, Hokfelt T, Dechelotte P (2008) Autoantibodies against appetite-regulating peptide hormones and neuropeptides: putative modulation by gut microflora. Nutrition 24(4): 348–359.
    1. Fetissov SO, Hamze Sinno M, Coquerel Q, Do Rego JC, Coeffier M, et al. (2008) Emerging role of autoantibodies against appetite-regulating neuropeptides in eating disorders. Nutrition 24(9): 854–859.
    1. Sekirov I, Russell SL, Antunes LC, Finlay BB (2010) Gut microbiota in health and disease. Physiol Rev 90(3): 859–904.
    1. Cani PD, Delzenne NM (2009) The role of the gut microbiota in energy metabolism and metabolic disease. Curr Pharm Des 15(13): 1546–1558.
    1. Routtenberg A, Kuznesof AW (1967) Self-starvation of rats living in activity wheels on a restricted feeding schedule. J Comp Physiol Psychol 64(3): 414–421.
    1. Kinzing KP, Hargrave SL (2010) Adolescent activity-based anorexia increases Anxiety-like behavior in Adulthood. Physiol Behav 101(2): 269–276.
    1. Seoane LM, Al-Massadi O, Barreiro F, Dieguez C, Casanueva F (2007) Growth hormone and somatostatin directly inhibit gastric ghrelin secretion. An in vitro organ culture system. J Endocrinol Invest 30(9): RC22–25.
    1. Seoane LM, Al-Massadi O, Caminos JE, Tovar SA, Dieguez C, et al. (2007) Sensory stimuli directly acting at the central nervous system regulate gastric ghrelin secretion. An ex vivo organ culture study. Endocrinology 148(8): 3998–4006.
    1. Queipo-Ortuño MI, Boto-Ordoñez M, Murri M, Gomez-Zumaquero JM, Clemente-Postigo M, et al. (2012) Influence of red wine polyphenols and ethanol on the gut microbiota ecology and biochemical biomarkers. Am J Clin Nutr 95(6): 1323–1334.
    1. Guo X, Xia X, Tang R, Zhou J, Zhao H, et al. (2008) Development of a real-time PCR method for Firmicutes and Bacteroidetes in faeces and its application to quantify intestinal population of obese and lean pigs. Lett Appl Microbiol 47(5): 367–373.
    1. Delroisse JM, Boulvin AL, Parmentier I, Dauphin RD, Vandenbol M (2008) Quantification of Bifidobacterium spp. and Lactobacillus spp. in rat fecal samples by real-time PCR. Microbiol Res 163(6): 663–670.
    1. Friswell MK, Gika H, Stratford IJ, Theodoridis G, Telfer B, et al. (2010) Site and strain-specific variation in gut microbiota profiles and metabolism in experimental mice. PLoS One 5(1): e8584.
    1. Stach JE, Maldonado LA, Ward AC, Goodfellow M, Bull AT (2003) New primers for the class Actinobacteria: application to marine and terrestrial environments. Environ Microbiol 5(10): 828–841.
    1. Matsuki T, Watanabe K, Fujimoto J, Takada T, Tanaka R (2004) Use of 16S rRNA gene-targeted group-specific primers for real-time PCR analysis of predominant bacteria in human feces. Appl Environ Microbiol 70(12): 7220–7228.
    1. Bekele AZ, Koike S, Kobayashi Y (2010) Genetic diversity and diet specificity of ruminal Prevotella revealed by 16S rRNA gene-based analysis. FEMS Microbiol Lett 305(1): 49–57.
    1. Rinttilä T, Kassinen A, Malinen E, Krogius L, Palva A (2004) Development of an extensive set of 16S rDNA-targeted primers for quantification of pathogenic and indigenous bacteria in faecal samples by real-time PCR. J Appl Microbiol 97(6): 1166–1177.
    1. Dridi B, Henry M, El Khéchine A, Raoult D, Drancourt M (2009) High prevalence of Methanobrevibacter smithii and Methanosphaera stadtmanae detected in the human gut using an improved DNA detection protocol. PLoS One 4(9): e7063.
    1. Lee ZMP, Bussema C, Schmidt TM (2009) rrnDB: documenting the number of rRNA and tRNA genes in bacteria and archaea. Nucleic Acids Res 37: D489–D493.
    1. Boakes RA, Juraskova I (2001) The role of drinking in the suppression of food intake by recent activity. Behav Neurosci 115(3): 718–730.
    1. Pardo M, Roca-Rivada A, Al-Massadi O, Seoane LM, Camiña JP, et al. (2010) Peripheral leptin and ghrelin receptors are regulated in a tissue-specific manner in activity-based anorexia. Peptides 31(10): 1912–1919.
    1. Ravussin Y, Koren O, Spor A, LeDuc C, Gutman R, et al. (2012) Responses of gut microbiota to diet composition and weight loss in lean and obese mice. Obesity 20(4): 738–747.
    1. Hebebrand J, Exner C, Hebebrand K, Holtkamp C, Casper RC, et al. (2003) Hyperactivity in patients with anorexia nervosa and in semistarved rats: evidence for a pivotal role of hypoleptinemia. Physiol Behav 79(1): 25–37.
    1. Giongo A, Gano KA, Crabb DB, Mukherjee N, Novelo LL, et al. (2011) Toward defining the autoimmune microbiome for type 1 diabetes. Isme J 5(1): 82–91.
    1. Armougom F, Henry M, Vialettes B, Raccah D, Raoult D (2009) Monitoring bacterial community of human gut microbiota reveals an increase in Lactobacillus in obese patients and Methanogens in anorexic patients. PLoS One 4(9): e7125.
    1. Deplancke B, Vidal O, Ganessunker D, Donovan SM, Mackie RI, et al. (2002) Selective growth of mucolytic bacteria including Clostridium perfringens in a neonatal piglet model of total parenteral nutrition. Am J Clin Nutr 76(5): 1117–1125.
    1. Miller RS, Hoskins LC (1981) Mucin degradation in human colon ecosystems. Fecal population densities of mucin-degrading bacteria estimated by a “most probable number” method. Gastroenterology 81(4): 759–765.
    1. Sonoyama K, Fujiwara R, Takemura N, Ogasawara T, Watanabe J, et al. (2009) Response of gut microbiota to fasting and hibernation in Syrian hamsters. Appl Environ Microbiol 75(20): 6451–6456.
    1. Burger-van Paassen N, Vincent A, Puiman PJ, van der Sluis M, Bouma J (2009) The regulation of intestinal mucin MUC2 expression by short-chain fatty acids: implications for epithelial protection. Biochem J 420(2): 211–219.
    1. Lin HV, Frassetto A, Kowalik EJ Jr, Nawrocki AR, Lu MM (2012) Butyrate and propionate protect against diet-induced obesity and regulate gut hormones via free fatty acid receptor 3-independent mechanisms. PLoS ONE 7(4): e35240.
    1. Ley RE (2010) Obesity and the human microbiome. Curr Opin Gastroenterol 26(1): 5–11.
    1. Turnbaugh PJ, Bäckhed F, Fulton L, Gordon JI (2008) Diet-induced obesity is linked to marked but reversible alterations in the mouse distal gut microbiome. Cell Host Microbe 3(4): 213–223.
    1. Turnbaugh PJ, Hamady M, Yatsunenko T, Cantarel BL, Duncan A, et al. (2009) A core gut microbiome in obese and lean twins. Nature 457(7228): 480–484.
    1. El Homsi M, Ducroc R, Claustre J, Jourdan G, Gertler A, et al. (2007) Leptin modulates the expression of secreted and membrane-associated mucins in colonic epithelial cells by targeting PKC, PI3K, and MAPK pathways. Am J Physiol Gastrointest Liver Physiol 293(1): 365–373.
    1. Plaisancie P, Ducroc R, El Homsi M, Tsocas A, Guilmeau S, et al. (2006) Luminal leptin activates mucinsecreting goblet cells in the large bowel. Am J Physiol Gastrointest Liver Physiol 290(4): 805–812.
    1. Brown CT, Davis-Richardson AG, Giongo A, Gano KA, Crabb DB, et al. (2011) Gut microbiome metagenomics analysis suggests a functional model for the development of autoimmunity for type 1 diabetes. PLoS One 6(10): e25792.
    1. Barcenilla A, Pryde SE, Martin JC, Duncan SH, Stewart CS, et al. (2000) Phylogenetic relationships of butyrate-producing bacteria from the human gut. Appl Environ Microbiol 66(4): 1654–1661.
    1. Matsumoto M, Inoue R, Tsukahara T, Ushida K, Chiji H, et al. (2008) Voluntary running exercise alters microbiota composition and increases n-butyrate concentration in the rat cecum. Biosci Biotechnol Biochem 72(2): 572–576.
    1. Lewis K, Lutgendorff F, Phan V, Soderholm JD, Sherman PM, et al. (2010) Enhanced Translocation of Bacteria Across Metabolically Stressed Epithelia is Reduced by Butyrate. Inflamm Bowel Dis 16(7): 1138–1148.
    1. Peng LY, Li Z, Green RS, Holzman IR, Lin J (2009) Butyrate enhances the intestinal barrier by facilitating tight junction assembly via activation of AMP-activated protein kinase in Caco-2 cell monolayers. J Nutr 139(9): 1619–1625.
    1. Hooda S, Vester Boler BM, Kerr KR, Dowd SE, Swanson KS (2012) The gut microbiome of kittens is affected by dietary protein: carbohydrate ratio and associated with blood metabolite and hormone concentrations. Br J Nutr 31: 1–10.

Source: PubMed

스폰서 및 공동 작업자

건강 상태

약물 개입

3
구독하다