Probiotic and Muscadine Grape Extract Interventions Shift the Gut Microbiome and Improve Metabolic Parameters in Female C57BL/6 Mice
<p>Dietary intervention modulates body condition and tissue morphology. (<b>A</b>) Body weight of female C57BL/6 mice following 13 weeks of exposure to diets and intervention strategies. (<b>B</b>) % change in body weight. (<b>C</b>) Visceral adipose tissue weight at study completion. (<b>D</b>) Normalized visceral adipose tissue weight at the end of the study. (<b>E</b>) Weight of the right lower (4/5) mammary gland at study completion. (<b>F</b>) Normalized inguinal mammary gland weight. <span class="html-italic">n</span> = 7–8. * <span class="html-italic">p</span> < 0.05, ** <span class="html-italic">p</span> < 0.01, *** <span class="html-italic">p</span> < 0.001, **** <span class="html-italic">p</span> < 0.0001.</p> "> Figure 2
<p>Diet composition and intervention strategies modulate gut microbiome composition. (<b>A</b>) Shannon diversity index of fecal samples collected after 13 weeks of diet and intervention exposure. (<b>B</b>) Principal component analysis of fecal microbial composition. (<b>C</b>) Proportional abundance of bacterial phyla identified in fecal samples. Each bar represents data collected from one mouse. (<b>D</b>) Proportional abundance of fecal Bacteroidetes. (<b>E</b>) Proportional abundance of fecal Firmicutes. (<b>F</b>) The ratio of Bacteroidetes to Firmicutes. <span class="html-italic">n</span> = 7–8, * <span class="html-italic">p</span> < 0.05.</p> "> Figure 3
<p>Dietary intake and MGE consumption mediate gut colonization of probiotic bacterial species. (<b>A</b>) Proportional abundance of bacterial species were identified in murine feces following 13 weeks of diet and intervention exposure. Each bar represents the fecal bacterial composition of a single mouse. (<b>B</b>–<b>E</b>) Proportional abundance of probiotic bacterial species identified in murine feces. (<b>B</b>) <span class="html-italic">Bifidobacterium</span>. (<b>C</b>) <span class="html-italic">Lactobacillus unclassified</span>. (<b>D</b><span class="html-italic">) Lactobacillus brevis</span>. (<b>E</b>) <span class="html-italic">Lactobacillus plantarum</span>. *** <span class="html-italic">p</span> < 0.001. **** <span class="html-italic">p</span> < 0.0001.</p> "> Figure 4
<p>Diet and intervention strategies mediate changes in visceral adipose tissue fibrosis. Representative 20× images of visceral adipose tissue stained with Picrosirius Red. Percentage of pixels positive for PicRed staining. * <span class="html-italic">p</span> < 0.05. ** <span class="html-italic">p</span> < 0.01. **** <span class="html-italic">p</span> < 0.0001.</p> "> Figure 5
<p>Diet and intervention strategies mediate changes in visceral adipose tissue immune cell infiltration. Representative 20× images of visceral adipose tissue stained with anti-F4/80. Number of F4/80-positive macrophages identified per million pixels. * <span class="html-italic">p</span> < 0.05. *** <span class="html-italic">p</span> < 0.001. **** <span class="html-italic">p</span> < 0.0001.</p> "> Figure 6
<p>Diet and intervention strategies mediate changes in visceral adipose tissue physiology and inflammation. (<b>A</b>) Representative 20× images of visceral adipose tissue stained with anti-MCP-1. (<b>B</b>) Percentage of pixels positive for anti-MCP-1 staining. (<b>C</b>) Average adipocyte diameter calculated from three representative adipocytes per image. * <span class="html-italic">p</span> < 0.05. ** <span class="html-italic">p</span> < 0.01. *** <span class="html-italic">p</span> < 0.001. **** <span class="html-italic">p</span> < 0.0001.</p> "> Figure 7
<p>Diet and intervention strategies mediate factors associated with intestinal inflammation. (<b>A</b>) Images of intestinal sections stained with H&E and Alcian Blue. (<b>B</b>) Average villus length measured in H&E images. (<b>C</b>) Alcian Blue-positive goblet cells counted per villus. * <span class="html-italic">p</span> < 0.05. ** <span class="html-italic">p</span> < 0.01. *** <span class="html-italic">p</span> < 0.001. **** <span class="html-italic">p</span> < 0.0001.</p> ">
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Animal Model
2.3. Gut Microbial Analysis
2.4. Immunohistochemistry
2.5. RT-PCR
2.6. Statistical Analysis
3. Results
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
- Rakhra, V.; Galappaththy, S.L.; Bulchandani, S.; Cabandugama, P.K. Obesity and the Western Diet: How We Got Here. Mol. Med. 2020, 117, 536–538. [Google Scholar]
- Christ, A.; Lauterbach, M.; Latz, E. Western Diet and the Immune System: An Inflammatory Connection. Immunity 2019, 51, 794–811. [Google Scholar] [CrossRef]
- Hosseini, B.; Saedisomeolia, A.; Wood, L.G.; Yaseri, M.; Tavasoli, S. Effects of pomegranate extract supplementation on inflammation in overweight and obese individuals: A randomized controlled clinical trial. Complement. Ther. Clin. Pract. 2016, 22, 44–50. [Google Scholar] [CrossRef] [PubMed]
- Lopez-Garcia, E.; Schulze, M.B.; Fung, T.T.; Meigs, J.B.; Rifai, N.; Manson, J.E.; Hu, F.B. Major dietary patterns are related to plasma concentrations of markers of inflammation and endothelial dysfunction. Am. J. Clin. Nutr. 2004, 80, 1029–1035. [Google Scholar] [CrossRef] [PubMed]
- Zinocker, M.K.; Lindseth, I.A. The Western Diet-Microbiome-Host Interaction and Its Role in Metabolic Disease. Nutrients 2018, 10, 365. [Google Scholar] [CrossRef] [PubMed]
- Suriano, F.; Nystrom, E.E.L.; Sergi, D.; Gustafsson, J.K. Diet, microbiota, and the mucus layer: The guardians of our health. Front. Immunol. 2022, 13, 953196. [Google Scholar] [CrossRef] [PubMed]
- Luis, A.S.; Hansson, G.C. Intestinal mucus and their glycans: A habitat for thriving microbiota. Cell Host Microbe 2023, 31, 1087–1100. [Google Scholar] [CrossRef] [PubMed]
- Groeger, D.; O’Mahony, L.; Murphy, E.F.; Bourke, J.F.; Dinan, T.G.; Kiely, B.; Shanahan, F.; Quigley, E.M. Bifidobacterium infantis 35624 modulates host inflammatory processes beyond the gut. Gut Microbes 2013, 4, 325–339. [Google Scholar] [CrossRef]
- Zocco, M.A.; dal Verme, L.Z.; Cremonini, F.; Piscaglia, A.C.; Nista, E.C.; Candelli, M.; Novi, M.; Rigante, D.; Cazzato, I.A.; Ojetti, V.; et al. Efficacy of Lactobacillus GG in maintaining remission of ulcerative colitis. Aliment. Pharmacol. Ther. 2006, 23, 1567–1574. [Google Scholar] [CrossRef]
- You, Q.; Chen, F.; Sharp, J.L.; Wang, X.; You, Y.; Zhang, C. High-performance liquid chromatography-mass spectrometry and evaporative light-scattering detector to compare phenolic profiles of muscadine grapes. J. Chromatogr. A 2012, 1240, 96–103. [Google Scholar] [CrossRef]
- Brown, J.C.; Huang, G.; Haley-Zitlin, V.; Jiang, X. Antibacterial effects of grape extracts on Helicobacter pylori. Appl. Environ. Microbiol. 2009, 75, 848–852. [Google Scholar] [CrossRef]
- Greenspan, P.; Bauer, J.D.; Pollock, S.H.; Gangemi, J.D.; Mayer, E.P.; Ghaffar, A.; Hargrove, J.L.; Hartle, D.K. Antiinflammatory properties of the muscadine grape (Vitis rotundifolia). J. Agric. Food Chem. 2005, 53, 8481–8484. [Google Scholar] [CrossRef] [PubMed]
- Ghanim, H.; Sia, C.L.; Korzeniewski, K.; Lohano, T.; Abuaysheh, S.; Marumganti, A.; Chaudhuri, A.; Dandona, P. A resveratrol and polyphenol preparation suppresses oxidative and inflammatory stress response to a high-fat, high-carbohydrate meal. J. Clin. Endocrinol. Metab. 2011, 96, 1409–1414. [Google Scholar] [CrossRef] [PubMed]
- Chappell, M.C.; Duncan, A.V.; Melo, A.C.; Schaich, C.L.; Pirro, N.T.; Diz, D.I.; Tallant, E.A.; Gallagher, P.E. Targeted UHPLC-MS Analysis Reveals Disparate Polyphenol Composition and Concentration in Muscadine Grape Supplements with Proportional Antioxidant Activity. Antioxidants 2022, 11, 2117. [Google Scholar] [CrossRef] [PubMed]
- Schloss, P.D.; Westcott, S.L.; Ryabin, T.; Hall, J.R.; Hartmann, M.; Hollister, E.B.; Lesniewski, R.A.; Oakley, B.B.; Parks, D.H.; Robinson, C.J.; et al. Introducing mothur: Open-source, platform-independent, community-supported software for describing and comparing microbial communities. Appl. Environ. Microbiol. 2009, 75, 7537–7541. [Google Scholar] [CrossRef]
- Krishnan, B.; Smith, T.L.; Dubey, P.; Zapadka, M.E.; Torti, F.M.; Willingham, M.C.; Tallant, E.A.; Gallagher, P.E. Angiotensin-(1-7) attenuates metastatic prostate cancer and reduces osteoclastogenesis. Prostate 2013, 73, 71–82. [Google Scholar] [CrossRef]
- Yahfoufi, N.; Alsadi, N.; Jambi, M.; Matar, C. The Immunomodulatory and Anti-Inflammatory Role of Polyphenols. Nutrients 2018, 10, 1618. [Google Scholar] [CrossRef]
- Cristofori, F.; Dargenio, V.N.; Dargenio, C.; Miniello, V.L.; Barone, M.; Francavilla, R. Anti-Inflammatory and Immunomodulatory Effects of Probiotics in Gut Inflammation: A Door to the Body. Front. Immunol. 2021, 12, 578386. [Google Scholar] [CrossRef]
- Marcelin, G.; Silveira, A.L.M.; Martins, L.B.; Ferreira, A.V.; Clement, K. Deciphering the cellular interplays underlying obesity-induced adipose tissue fibrosis. J. Clin. Investig. 2019, 129, 4032–4040. [Google Scholar] [CrossRef]
- Gourineni, V.; Shay, N.F.; Chung, S.; Sandhu, A.K.; Gu, L. Muscadine grape (Vitis rotundifolia) and wine phytochemicals prevented obesity-associated metabolic complications in C57BL/6J mice. J. Agric. Food Chem. 2012, 60, 7674–7681. [Google Scholar] [CrossRef]
- Borgeraas, H.; Johnson, L.K.; Skattebu, J.; Hertel, J.K.; Hjelmesaeth, J. Effects of probiotics on body weight, body mass index, fat mass and fat percentage in subjects with overweight or obesity: A systematic review and meta-analysis of randomized controlled trials. Obes. Rev. 2018, 19, 219–232. [Google Scholar] [CrossRef] [PubMed]
- Sanchez, M.; Darimont, C.; Drapeau, V.; Emady-Azar, S.; Lepage, M.; Rezzonico, E.; Ngom-Bru, C.; Berger, B.; Philippe, L.; Ammon-Zuffrey, C.; et al. Effect of Lactobacillus rhamnosus CGMCC1.3724 supplementation on weight loss and maintenance in obese men and women. Br. J. Nutr. 2014, 111, 1507–1519. [Google Scholar] [CrossRef] [PubMed]
- Suarez-Zamorano, N.; Fabbiano, S.; Chevalier, C.; Stojanovic, O.; Colin, D.J.; Stevanovic, A.; Veyrat-Durebex, C.; Tarallo, V.; Rigo, D.; Germain, S.; et al. Microbiota depletion promotes browning of white adipose tissue and reduces obesity. Nat. Med. 2015, 21, 1497–1501. [Google Scholar] [CrossRef] [PubMed]
- Newman, T.M.; Shively, C.A.; Register, T.C.; Appt, S.E.; Yadav, H.; Colwell, R.R.; Fanelli, B.; Dadlani, M.; Graubics, K.; Nguyen, U.T.; et al. Diet, obesity, and the gut microbiome as determinants modulating metabolic outcomes in a non-human primate model. Microbiome 2021, 9, 100. [Google Scholar] [CrossRef] [PubMed]
- Lozupone, C.A.; Stombaugh, J.I.; Gordon, J.I.; Jansson, J.K.; Knight, R. Diversity, stability and resilience of the human gut microbiota. Nature 2012, 489, 220–230. [Google Scholar] [CrossRef] [PubMed]
- Ley, R.E. Obesity and the human microbiome. Curr. Opin. Gastroenterol. 2010, 26, 5–11. [Google Scholar] [CrossRef]
- Zhao, L.; Zhang, Q.; Ma, W.; Tian, F.; Shen, H.; Zhou, M. A combination of quercetin and resveratrol reduces obesity in high-fat diet-fed rats by modulation of gut microbiota. Food Funct. 2017, 8, 4644–4656. [Google Scholar] [CrossRef]
- Stojanov, S.; Berlec, A.; Strukelj, B. The Influence of Probiotics on the Firmicutes/Bacteroidetes Ratio in the Treatment of Obesity and Inflammatory Bowel disease. Microorganisms 2020, 8, 1715. [Google Scholar] [CrossRef]
- Turnbaugh, P.J.; Ley, R.E.; Mahowald, M.A.; Magrini, V.; Mardis, E.R.; Gordon, J.I. An obesity-associated gut microbiome with increased capacity for energy harvest. Nature 2006, 444, 1027–1031. [Google Scholar] [CrossRef]
- Xue, B.; Xie, J.; Huang, J.; Chen, L.; Gao, L.; Ou, S.; Wang, Y.; Peng, X. Plant polyphenols alter a pathway of energy metabolism by inhibiting fecal Bacteroidetes and Firmicutes in vitro. Food Funct. 2016, 7, 1501–1507. [Google Scholar] [CrossRef]
- Tabasco, R.; Sanchez-Patan, F.; Monagas, M.; Bartolome, B.; Victoria Moreno-Arribas, M.; Pelaez, C.; Requena, T. Effect of grape polyphenols on lactic acid bacteria and bifidobacteria growth: Resistance and metabolism. Food Microbiol. 2011, 28, 1345–1352. [Google Scholar] [CrossRef]
- Pellegrinelli, V.; Heuvingh, J.; du Roure, O.; Rouault, C.; Devulder, A.; Klein, C.; Lacasa, M.; Clement, E.; Lacasa, D.; Clement, K. Human adipocyte function is impacted by mechanical cues. J. Pathol. 2014, 233, 183–195. [Google Scholar] [CrossRef]
- Hotamisligil, G.S.; Shargill, N.S.; Spiegelman, B.M. Adipose expression of tumor necrosis factor-alpha: Direct role in obesity-linked insulin resistance. Science 1993, 259, 87–91. [Google Scholar] [CrossRef] [PubMed]
- Suriano, F.; Vieira-Silva, S.; Falony, G.; Roumain, M.; Paquot, A.; Pelicaen, R.; Regnier, M.; Delzenne, N.M.; Raes, J.; Muccioli, G.G.; et al. Novel insights into the genetically obese (ob/ob) and diabetic (db/db) mice: Two sides of the same coin. Microbiome 2021, 9, 147. [Google Scholar] [CrossRef] [PubMed]
- Weisberg, S.P.; McCann, D.; Desai, M.; Rosenbaum, M.; Leibel, R.L.; Ferrante, A.W., Jr. Obesity is associated with macrophage accumulation in adipose tissue. J. Clin. Investig. 2003, 112, 1796–1808. [Google Scholar] [CrossRef]
- Kanda, H.; Tateya, S.; Tamori, Y.; Kotani, K.; Hiasa, K.; Kitazawa, R.; Kitazawa, S.; Miyachi, H.; Maeda, S.; Egashira, K.; et al. MCP-1 contributes to macrophage infiltration into adipose tissue, insulin resistance, and hepatic steatosis in obesity. J. Clin. Investig. 2006, 116, 1494–1505. [Google Scholar] [CrossRef] [PubMed]
- Belkaid, Y.; Hand, T.W. Role of the microbiota in immunity and inflammation. Cell 2014, 157, 121–141. [Google Scholar] [CrossRef]
- Lv, X.C.; Chen, M.; Huang, Z.R.; Guo, W.L.; Ai, L.Z.; Bai, W.D.; Yu, X.D.; Liu, Y.L.; Rao, P.F.; Ni, L. Potential mechanisms underlying the ameliorative effect of Lactobacillus paracasei FZU103 on the lipid metabolism in hyperlipidemic mice fed a high-fat diet. Food Res. Int. 2021, 139, 109956. [Google Scholar] [CrossRef]
- Kowalska, K.; Dembczynski, R.; Golabek, A.; Olkowicz, M.; Olejnik, A. ROS Modulating Effects of Lingonberry (Vaccinium vitis-idaea L.) Polyphenols on Obese Adipocyte Hypertrophy and Vascular Endothelial Dysfunction. Nutrients 2021, 13, 885. [Google Scholar] [CrossRef]
- Coltman, C.E.; Steele, J.R.; McGhee, D.E. Breast volume is affected by body mass index but not age. Ergonomics 2017, 60, 1576–1585. [Google Scholar] [CrossRef]
- Wu, J.; Crowe, D.L. Molecular and cellular basis of mammary gland fibrosis and cancer risk. Int. J. Cancer 2019, 144, 2239–2253. [Google Scholar] [CrossRef] [PubMed]
- Brady, N.J.; Chuntova, P.; Schwertfeger, K.L. Macrophages: Regulators of the Inflammatory Microenvironment during Mammary Gland Development and Breast Cancer. Mediat. Inflamm. 2016, 2016, 4549676. [Google Scholar] [CrossRef] [PubMed]
- Dawson, C.A.; Pal, B.; Vaillant, F.; Gandolfo, L.C.; Liu, Z.; Bleriot, C.; Ginhoux, F.; Smyth, G.K.; Lindeman, G.J.; Mueller, S.N.; et al. Tissue-resident ductal macrophages survey the mammary epithelium and facilitate tissue remodelling. Nat. Cell Biol. 2020, 22, 546–558. [Google Scholar] [CrossRef]
- Sun, X.; Glynn, D.J.; Hodson, L.J.; Huo, C.; Britt, K.; Thompson, E.W.; Woolford, L.; Evdokiou, A.; Pollard, J.W.; Robertson, S.A.; et al. CCL2-driven inflammation increases mammary gland stromal density and cancer susceptibility in a transgenic mouse model. Breast Cancer Res. 2017, 19, 4. [Google Scholar] [CrossRef] [PubMed]
- Parker, A.; Vaux, L.; Patterson, A.M.; Modasia, A.; Muraro, D.; Fletcher, A.G.; Byrne, H.M.; Maini, P.K.; Watson, A.J.M.; Pin, C. Elevated apoptosis impairs epithelial cell turnover and shortens villi in TNF-driven intestinal inflammation. Cell Death Dis. 2019, 10, 108. [Google Scholar] [CrossRef] [PubMed]
- Tanovic, A.; Fernandez, E.; Jimenez, M. Alterations in intestinal contractility during inflammation are caused by both smooth muscle damage and specific receptor-mediated mechanisms. Croat. Med. J. 2006, 47, 318–326. [Google Scholar] [PubMed]
- Wu, Z.; Huang, S.; Li, T.; Li, N.; Han, D.; Zhang, B.; Xu, Z.Z.; Zhang, S.; Pang, J.; Wang, S.; et al. Gut microbiota from gr een tea polyphenol-dosed mice improves intestinal epithelial homeostasis and ameliorates experimental colitis. Microbiome 2021, 9, 184. [Google Scholar] [CrossRef] [PubMed]
- Di Giacinto, C.; Marinaro, M.; Sanchez, M.; Strober, W.; Boirivant, M. Probiotics ameliorate recurrent Th1-mediated murine colitis by inducing IL-10 and IL-10-dependent TGF-beta-bearing regulatory cells. J. Immunol. 2005, 174, 3237–3246. [Google Scholar] [CrossRef]
- Kim, Y.S.; Ho, S.B. Intestinal goblet cells and mucins in health and disease: Recent insights and progress. Curr. Gastroenterol. Rep. 2010, 12, 319–330. [Google Scholar] [CrossRef]
- Deplancke, B.; Gaskins, H.R. Microbial modulation of innate defense: Goblet cells and the intestinal mucus layer. Am. J. Clin. Nutr. 2001, 73, 1131S–1141S. [Google Scholar] [CrossRef]
CD | WD | |
---|---|---|
(TD.08806) | (TD.180300) | |
Protein (% kcal) | 20.5% | 15.9% |
Carbohydrates (% kcal) | 69.1% | 39.6% |
Fat (% kcal) | 10% | 44.5% |
Saturated Fat | 27% | 43.3% |
Monounsaturated Fat | 36.5% | 35.1% |
Polyunsaturated Fat | 36.5% | 20.5% |
Sucrose | 11.2% | 25.5% |
Cholesterol (mg/kg) | 0.017 | 0.056 |
Sodium (g/kg) | 0.28 | 1.59 |
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content. |
© 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Newman, T.M.; Wilson, A.S.; Clear, K.Y.J.; Tallant, E.A.; Gallagher, P.E.; Cook, K.L. Probiotic and Muscadine Grape Extract Interventions Shift the Gut Microbiome and Improve Metabolic Parameters in Female C57BL/6 Mice. Cells 2023, 12, 2599. https://doi.org/10.3390/cells12222599
Newman TM, Wilson AS, Clear KYJ, Tallant EA, Gallagher PE, Cook KL. Probiotic and Muscadine Grape Extract Interventions Shift the Gut Microbiome and Improve Metabolic Parameters in Female C57BL/6 Mice. Cells. 2023; 12(22):2599. https://doi.org/10.3390/cells12222599
Chicago/Turabian StyleNewman, Tiffany M., Adam S. Wilson, Kenysha Y. J. Clear, E. Ann Tallant, Patricia E. Gallagher, and Katherine L. Cook. 2023. "Probiotic and Muscadine Grape Extract Interventions Shift the Gut Microbiome and Improve Metabolic Parameters in Female C57BL/6 Mice" Cells 12, no. 22: 2599. https://doi.org/10.3390/cells12222599