Intestinal nerve cell injury occurs prior to insulin resistance in female mice ingesting a high-fat diet
Diabetic patients suffer from gastrointestinal disorders associated with dysmotility, enteric neuropathy and dysbiosis of gut microbiota; however, gender differences are not fully known. Previous studies have shown that a high-fat diet (HFD) causes type two diabetes (T2D) in male mice after 4–8 weeks but only does so in female mice after 16 weeks. This study seeks to determine whether sex influences the development of intestinal dysmotility, enteric neuropathy and dysbiosis in mice fed HFD. We fed 8-week-old C57BL6 male and female mice a standard chow diet (SCD) or a 72% kcal HFD for 8 weeks. We analyzed the associations between sex and intestinal dysmotility, neuropathy and dysbiosis using motility assays, immunohistochemistry and next-generation sequencing. HFD ingestion caused obesity, glucose intolerance and insulin resistance in male but not female mice. However, HFD ingestion slowed intestinal propulsive motility in both male and female mice. This was associated with decreased inhibitory neuromuscular transmission, loss of myenteric inhibitory motor neurons and axonal swelling and loss of cytoskeletal filaments. HFD induced dysbiosis and changed the abundance of specific bacteria, especially Allobaculum, Bifidobacterium and Lactobacillus, which correlated with dysmotility and neuropathy. Female mice had higher immunoreactivity and numbers of myenteric inhibitory motor neurons, matching larger amplitudes of inhibitory junction potentials. This study suggests that sex influences the development of HFD-induced metabolic syndrome but dysmotility, neuropathy and dysbiosis occur independent of sex and prior to T2D conditions. Gastrointestinal dysmotility, neuropathy and dysbiosis might play a crucial role in the pathophysiology of T2D in humans irrespective of sex.
KeywordsGut microbiota Diabetes mellitus Gastrointestinal motility Enteric nervous system Diabetic enteric neuropathy
We would like to thank Catherine Brands and the UI Laboratory Animal Research Facility staff for their assistance during animal handling and Forrest Potter and Ann Norton (IBEST Optical Imaging Core) for their assistance with imaging and analysis. We thank Dan New, Dr. Matt Settles, Dr. Celeste Brown, Dr. Ben Ridenhour and the IBEST Genomics Resources Core for their help with microbial community analysis. We also thank Dr. Vanda A. Lennon of the Mayo Clinic for the gift of ANNA1 positive human serum.
Conception and design: YN, LF and OB. Development of methodology: YN, MG and OB. Acquisition of data: YN, RE, KS, CM, HE, KH, HO, JM, GM, MG and OB. Analysis and interpretation of data: YN, MG and OB. Writing, review and/or revision of the manuscript: YN, LF, MG and OB. Study supervision: OB.
The research reported in this publication was supported by the University of Idaho—Dyess Faculty Fellowship and Institutional Development Awards (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant numbers P30GM103324 and P20 GM103408.
Compliance with ethical standards
The authors declare that they have no conflict of interest.
All authors have no competing interests financial or otherwise and have nothing to disclose.
All applicable international, national and/or institutional guidelines for the care and use of animals were followed and all procedures performed in studies involving animals were in accordance with the ethical standards of the University of Idaho.
All authors declare that this research was done by strictly adhering to the rules of good scientific practice and are responsible for its content. All experiments were performed in a manner that maximized rigor and reproducibility and without bias.
- Abrahamsson H (1995) Gastrointestinal motility disorders in patients with diabetes mellitus. J Intern Med 237:403–409. https://doi.org/10.1111/j.1365-2796.1995.tb01194.x CrossRefGoogle Scholar
- Balemba OB, Bhattarai Y, Stenkamp-Strahm C, Lesakit MSB, Mawe GM (2010) The traditional antidiarrheal remedy, Garcinia buchananii stem bark extract, inhibits propulsive motility and fast synaptic potentials in the guinea pig distal colon. Neurogastroenterol Motil 22:1332–1339. https://doi.org/10.1111/j.1365-2982.2010.01583.x CrossRefGoogle Scholar
- Bhattarai Y, Fried D, Gulbransen B, Kadrofske M, Fernandes R, Xu H, Galligan J (2016) High-fat diet-induced obesity alters nitric oxide-mediated neuromuscular transmission and smooth muscle excitability in the mouse distal colon. Am J Physiol Gastrointest Liver Physiol 311:G210–G220. https://doi.org/10.1152/ajpgi.00085.2016 CrossRefGoogle Scholar
- Boulangé CL, Neves AL, Chilloux J, Nicholson JK, Dumas M-E (2016) Impact of the gut microbiota on inflammation, obesity, and metabolic disease. Genome Med 8(42). https://doi.org/10.1186/s13073-016-0303-2
- Cani PD, Amar J, Iglesias MA, Poggi M, Knauf C, Bastelica D, Neyrinck AM, Fava F, Tuohy KM, Chabo C, Waget A, Delmee E, Cousin B, Sulpice T, Chamontin B, Ferrieres J, Tanti J-FJ-F, Gibson GR, Casteilla L, Delzenne NM, Alessi MC, Burcelin R, Delmée E, Cousin B, Sulpice T, Chamontin B, Ferrières J, Tanti J-FJ-F, Gibson GR, Casteilla L, Delzenne NM, Alessi MC, Burcelin R (2007) Metabolic endotoxemia initiates obesity and insulin resistance. Diabetes 56:1761–1772. https://doi.org/10.2337/db06-1491 CrossRefGoogle Scholar
- Centers for Disease Control and Prevention (2017) National Diabetes Statistics Report, 2017 Estimates of diabetes and its burden in the epidemiologic estimation methods. https://www.cdc.gov/diabetes/data/statistics/statistics-report.html
- Chandrasekharan B, Anitha M, Blatt R, Shahnavaz N, Kooby D, Staley C, Mwangi S, Jones DP, Sitaraman SV, Srinivasan S (2011) Colonic motor dysfunction in human diabetes is associated with enteric neuronal loss and increased oxidative stress. Neurogastroenterol Motil 23:131–e26. https://doi.org/10.1111/j.1365-2982.2010.01611.x CrossRefGoogle Scholar
- Everard A, Lazarevic V, Gaïa N, Johansson M, Ståhlman M, Backhed F, Delzenne NM, Schrenzel J, François P, Cani PD (2014) Microbiome of prebiotic-treated mice reveals novel targets involved in host response during obesity. ISME J. https://doi.org/10.1038/ismej.2014.45
- Grenham S, Clarke G, Cryan JF, Dinan TG (2011) Brain-gut-microbe communication in health and disease. Front Physiol 2(94). https://doi.org/10.3389/fphys.2011.00094
- Grover M, Farrugia G, Lurken MS, Bernard CE, Faussone-Pellegrini MS, Smyrk TC, Parkman HP, Abell TL, Snape WJ, Hasler WL, Ünalp-Arida A, Nguyen L, Koch KL, Calles J, Lee L, Tonascia J, Hamilton FA, Pasricha PJ, NIDDK Gastroparesis Clinical Research Consortium (2011) Cellular changes in diabetic and idiopathic gastroparesis. Gastroenterology 140:1575–85.e8. https://doi.org/10.1053/j.gastro.2011.01.046 CrossRefGoogle Scholar
- Haro C, Rangel-Zúñiga OA, Alcalá-Díaz JF, Gómez-Delgado F, Pérez-Martínez P, Delgado-Lista J, Quintana-Navarro GM, Landa BB, Navas-Cortés JA, Tena-Sempere M, Clemente JC, López-Miranda J, Pérez-Jiménez F, Camargo A (2016) Intestinal microbiota is influenced by gender and body mass index. PLoS One 11:e0154090. https://doi.org/10.1371/journal.pone.0154090 CrossRefGoogle Scholar
- Hoffman JM, Brooks EM, Mawe GM (2010) Gastrointestinal Motility Monitor (GIMM). J Vis Exp. https://doi.org/10.3791/2435
- Kashyap PC, Marcobal A, Ursell LK, Larauche M, Duboc H, Earle KA, Sonnenburg ED, Ferreyra JA, Higginbottom SK, Million M, Tache Y, Pasricha PJ, Knight R, Farrugia G, Sonnenburg JL (2013) Complex interactions among diet, gastrointestinal transit, and gut microbiota in humanized mice. Gastroenterology 144:967–977. https://doi.org/10.1053/j.gastro.2013.01.047 CrossRefGoogle Scholar
- Kearney PM, Whelton M, Reynolds K, Whelton PK, He J (2004) Worldwide prevalence of hypertension: a systematic review. J Hypertens 22:11–19Google Scholar
- Morselli E, Fuente-Martin E, Finan B, Kim M, Frank A, Garcia-Caceres C, Navas CR, Gordillo R, Neinast M, Kalainayakan SP, Li DL, Gao Y, Yi C-X, Hahner L, Palmer BF, Tschöp MH, Clegg DJ (2014) Hypothalamic PGC-1α protects against high-fat diet exposure by regulating ERα. Cell Rep 9:633–645. https://doi.org/10.1016/j.celrep.2014.09.025 CrossRefGoogle Scholar
- Pasricha PJ, Pehlivanov ND, Gomez G, Vittal H, Lurken MS, Farrugia G (2008) Changes in the gastric enteric nervous system and muscle: a case report on two patients with diabetic gastroparesis. BMC Gastroenterol 8(21). https://doi.org/10.1186/1471-230X-8-21
- Payne AN, Chassard C, Zimmermann M, Müller P, Stinca S, Lacroix C (2011) The metabolic activity of gut microbiota in obese children is increased compared with normal-weight children and exhibits more exhaustive substrate utilization. Nutr Diabetes 1:e12. https://doi.org/10.1038/nutd.2011.8 CrossRefGoogle Scholar
- Ravel J, Gajer P, Abdo Z, Schneider GM, Koenig SSK, McCulle SL, Karlebach S, Gorle R, Russell J, Tacket CO, Brotman RM, Davis CC, Ault K, Peralta L, Forney LJ (2011) Vaginal microbiome of reproductive-age women. Proc Natl Acad Sci U S A 108(Suppl):4680–4687. https://doi.org/10.1073/pnas.1002611107 CrossRefGoogle Scholar
- Reichardt F, Chassaing B, Nezami BG, Li G, Tabatabavakili S, Mwangi S, Uppal K, Liang B, Vijay-Kumar M, Jones D, Gewirtz AT, Srinivasan S (2017) Western diet induces colonic nitrergic myenteric neuropathy and dysmotility in mice via saturated fatty acid- and lipopolysaccharide-induced TLR4 signalling. J Physiol 595:1831–1846. https://doi.org/10.1113/JP273269 CrossRefGoogle Scholar
- Rivera LR, Leung C, Pustovit RV, Hunne BL, Andrikopoulos S, Herath C, Testro A, Angus PW, Furness JB (2014) Damage to enteric neurons occurs in mice that develop fatty liver disease but not diabetes in response to a high-fat diet. Neurogastroenterol Motil 26:1188–1199. https://doi.org/10.1111/nmo.12385 CrossRefGoogle Scholar
- Spencer NJ, Hennig GW, Smith TK (2001) Spatial and temporal coordination of junction potentials in circular muscle of guinea-pig distal colon. J Physiol 535:565–578. https://doi.org/10.1111/j.1469-7793.2001.00565.x
- Stenkamp-Strahm CM, Nyavor YEA, Kappmeyer AJ, Horton S, Gericke M, Balemba OB (2015) Prolonged high fat diet ingestion, obesity, and type 2 diabetes symptoms correlate with phenotypic plasticity in myenteric neurons and nerve damage in the mouse duodenum. Cell Tissue Res 361:411–426. https://doi.org/10.1007/s00441-015-2132-9 CrossRefGoogle Scholar
- Ussar S, Griffin NW, Bezy O, Fujisaka S, Vienberg S, Softic S, Deng L, Bry L, Gordon JI, Kahn CR (2015) Interactions between gut microbiota, host genetics and diet modulate the predisposition to obesity and metabolic syndrome. Cell Metab 22:516–530. https://doi.org/10.1016/j.cmet.2015.07.007 CrossRefGoogle Scholar
- Van Hul M, Geurts L, Plovier H, Druart C, Everard A, Ståhlman M, Rhimi M, Chira K, Teissedre P-L, Delzenne NM, Maguin E, Guilbot A, Brochot A, Gerard P, Bäckhed F, Cani PD (2017) Reduced obesity, diabetes and steatosis upon cinnamon and grape pomace are associated with changes in gut microbiota and markers of gut barrier. Am J Physiol Endocrinol Metab 314:ajpendo.00107.2017. https://doi.org/10.1152/ajpendo.00107.2017 Google Scholar
- Wu RY, Pasyk M, Wang B, Forsythe P, Bienenstock J, Mao Y-K, Sharma P, Stanisz AM, Kunze WA (2013) Spatiotemporal maps reveal regional differences in the effects on gut motility for Lactobacillus reuteri and rhamnosus strains. Neurogastroenterol Motil. https://doi.org/10.1111/nmo.12072