Abstract
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.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
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
Anitha M, Gondha C, Sutliff R, Parsadanian A, Mwangi S, Sitaraman SV, Srinivasan S (2006) GDNF rescues hyperglycemia-induced diabetic enteric neuropathy through activation of the PI3K/Akt pathway. J Clin Invest 116:344–356. https://doi.org/10.1172/JCI26295
Aziz Q, Doré J, Emmanuel A, Guarner F, Quigley EMM (2013) Gut microbiota and gastrointestinal health: current concepts and future directions. Neurogastroenterol Motil 25:4–15. https://doi.org/10.1111/nmo.12046
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
Bertrand RL, Senadheera S, Markus I, Liu L, Howitt L, Chen H, Murphy TV, Sandow SL, Bertrand PP (2011) A Western diet increases serotonin availability in rat small intestine. Endocrinology 152:36–47. https://doi.org/10.1210/en.2010-0377
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
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
Bridgewater LC, Zhang C, Wu Y, Hu W, Zhang Q, Wang J, Li S, Zhao L (2017) Gender-based differences in host behavior and gut microbiota composition in response to high fat diet and stress in a mouse model. Sci Rep 7:10776. https://doi.org/10.1038/s41598-017-11069-4
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
Cefalu WT (2006) Animal models of type 2 diabetes: clinical presentation and pathophysiological relevance to the human condition. ILAR J 47:186–198. https://doi.org/10.1093/ilar.47.3.186
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, Srinivasan S (2007) Diabetes and the enteric nervous system. Neurogastroenterol Motil 19:951–960. https://doi.org/10.1111/j.1365-2982.2007.01023.x
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
Collins J, Borojevic R, Verdu EF, Huizinga JD, Ratcliffe EM (2014) Intestinal microbiota influence the early postnatal development of the enteric nervous system. Neurogastroenterol Motil 26:98–107. https://doi.org/10.1111/nmo.12236
Degen LP, Phillips SF (1996) Variability of gastrointestinal transit in healthy women and men. Gut 39:299–305. https://doi.org/10.1136/gut.39.2.299
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
Farrugia G (2015) Histologic changes in diabetic gastroparesis. Gastroenterol Clin N Am 44:31–38. https://doi.org/10.1016/j.gtc.2014.11.004
Ford ES (2005) Prevalence of the metabolic syndrome defined by the International Diabetes Federation among adults in the U.S. Diabetes Care 28:2745–2749
France M, Skorich E, Kadrofske M, Swain GM, Galligan JJ (2016) Sex-related differences in small intestinal transit and serotonin dynamics in high-fat-diet-induced obesity in mice. Exp Physiol 101:81–99. https://doi.org/10.1113/EP085427
Fu X-Y, Li Z, Zhang N, Yu H-T, Wang S-R, Liu J-R (2014) Effects of gastrointestinal motility on obesity. Nutr Metab (Lond) 11:3. https://doi.org/10.1186/1743-7075-11-3
Furness JB (2012) The enteric nervous system and neurogastroenterology. Nat Rev Gastroenterol Hepatol 9:286–294. https://doi.org/10.1038/nrgastro.2012.32
Greetham HL, Gibson GR, Giffard C, Hippe H, Merkhoffer B, Steiner U, Falsen E, Collins MD (2004) Allobaculum stercoricanis gen. nov., sp. nov., isolated from canine feces. Anaerobe 10:301–307. https://doi.org/10.1016/j.anaerobe.2004.06.004
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
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
Hoffman JM, Brooks EM, Mawe GM (2010) Gastrointestinal Motility Monitor (GIMM). J Vis Exp. https://doi.org/10.3791/2435
Horváth VJ, Vittal H, Ordög T (2005) Reduced insulin and IGF-I signaling, not hyperglycemia, underlies the diabetes-associated depletion of interstitial cells of Cajal in the murine stomach. Diabetes 54:1528–1533. https://doi.org/10.2337/diabetes.54.5.1528
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
Kearney PM, Whelton M, Reynolds K, Whelton PK, He J (2004) Worldwide prevalence of hypertension: a systematic review. J Hypertens 22:11–19
Kunze WA, Furness JB (1999) The enteric nervous system and regulation of intestinal motility. Annu Rev Physiol 61:117–142. https://doi.org/10.1146/annurev.physiol.61.1.117
Lei B, Mace B, Dawson HN, Warner DS, Laskowitz DT, James ML (2014) Anti-inflammatory effects of progesterone in lipopolysaccharide-stimulated BV-2 microglia. PLoS One 9:e103969. https://doi.org/10.1371/journal.pone.0103969
Li H, Qi T, sen HZ, Ying Y, Zhang Y, Wang B, Ye L, Zhang B, ling CD, Chen J (2017) Relationship between gut microbiota and type 2 diabetic erectile dysfunction in Sprague-Dawley rats. J Huazhong Univ Sci Technol - Med Sci 37:523–530. https://doi.org/10.1007/s11596-017-1767-z
Matthews DR, Hosker JP, Rudenski AS, Naylor BA, Treacher DF, Turner RC (1985) Homeostasis model assessment: insulin resistance and β-cell function from fasting plasma glucose and insulin concentrations in man. Diabetologia 28:412–419. https://doi.org/10.1007/BF00280883
Meleine M, Matricon J (2014) Gender-related differences in irritable bowel syndrome: potential mechanisms of sex hormones. World J Gastroenterol 20:6725–6743. https://doi.org/10.3748/wjg.v20.i22.6725
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
Nyavor YEA, Balemba OB (2017) Diet-induced dysmotility and neuropathy in the gut precedes endotoxaemia and metabolic syndrome: the chicken and the egg revisited. J Physiol 595:1441–1442. https://doi.org/10.1113/JP273888
Ozbey N, Sencer E, Molvalilar S, Orhan Y (2002) Body fat distribution and cardiovascular disease risk factors in pre- and postmenopausal obese women with similar BMI. Endocr J 49:503–509
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
Pettersson US, Waldén TB, Carlsson PO, Jansson L, Phillipson M (2012) Female mice are protected against high-fat diet induced metabolic syndrome and increase the regulatory T cell population in adipose tissue. PLoS One 7:e46057. https://doi.org/10.1371/journal.pone.0046057
Quigley EMM (2011) Microflora modulation of motility. J Neurogastroenterol Motil 17:140–147. https://doi.org/10.5056/jnm.2011.17.2.140
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
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
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
Roberts JA, Durnin L, Sharkey KA, Mutafova-Yambolieva VN, Mawe GM (2013) Oxidative stress disrupts purinergic neuromuscular transmission in the inflamed colon. J Physiol 591:3725–3737. https://doi.org/10.1113/jphysiol.2013.254136
Sandireddy R, Yerra VG, Areti A, Komirishetty P, Kumar A (2014) Neuroinflammation and oxidative stress in diabetic neuropathy: futuristic strategies based on these targets. Int J Endocrinol 2014:674987. https://doi.org/10.1155/2014/674987
Smyth S, Heron A (2006) Diabetes and obesity: the twin epidemics. Nat Med 12:75–80. https://doi.org/10.1038/nm0106-75
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, Kappmeyer AJ, Schmalz JT, Gericke M, Balemba O (2013) High-fat diet ingestion correlates with neuropathy in the duodenum myenteric plexus of obese mice with symptoms of type 2 diabetes. Cell Tissue Res 354:381–394. https://doi.org/10.1007/s00441-013-1681-z
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
Sugiyama MG, Agellon LB (2012) Sex differences in lipid metabolism and metabolic disease risk. Biochem Cell Biol 90:124–141. https://doi.org/10.1139/o11-067
Tachon S, Zhou J, Keenan M, Martin R, Marco ML (2013) The intestinal microbiota in aged mice is modulated by dietary resistant starch and correlated with improvements in host responses. FEMS Microbiol Ecol 83:299–309. https://doi.org/10.1111/j.1574-6941.2012.01475.x
Turnbaugh PJ, Ley RE, Mahowald MA, Magrini V, Mardis ER, Gordon JI (2006) An obesity-associated gut microbiome with increased capacity for energy harvest. Nature 444:1027–1031. https://doi.org/10.1038/nature05414
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
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
Vincent AM, Russell JW, Low P, Feldman EL (2004) Oxidative stress in the pathogenesis of diabetic neuropathy. Endocr Rev 25:612–628. https://doi.org/10.1210/er.2003-0019
Voss U, Sand E, Olde B, Ekblad E, Donath M (2013) Enteric neuropathy can be induced by high fat diet in vivo and palmitic acid exposure in vitro. PLoS One 8:e81413. https://doi.org/10.1371/journal.pone.0081413
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
Yarandi SS, Srinivasan S (2014) Diabetic gastrointestinal motility disorders and the role of enteric nervous system: current status and future directions. Neurogastroenterol Motil 26:611–624. https://doi.org/10.1111/nmo.12330
Yuan S, Cohen DB, Ravel J, Abdo Z, Forney LJ (2012) Evaluation of methods for the extraction and purification of DNA from the human microbiome. PLoS One 7:e33865. https://doi.org/10.1371/journal.pone.0033865
Acknowledgements
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.
Funding
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.
Author information
Authors and Affiliations
Contributions
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.
Corresponding author
Ethics declarations
Competing interests
The authors declare that they have no conflict of interest.
Disclosures
All authors have no competing interests financial or otherwise and have nothing to disclose.
Ethical approval
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.
Ethical statement
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.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
ESM 1
(DOCX 107 kb)
Supplementary Fig. 1
HFD has different effects on the metabolism of male and female mice. Serum concentrations of leptin (a), resistin (b) and plasmogen activator inhibitor 1 (c, PAI-1) were measured by Milliplex assay. HFD increased the leptin concentration in both male and female mice but male mice had a significantly higher leptin concentration than female mice overall (a). Conversely, resistin was not affected by HFD in female mice even though it was increased in male mice (b). PAI-1 concentrations were increased by HFD ingestion in male mice but unaffected (reduced) in female mice (c). Data are expressed as mean ± SEM and analyzed by one-way analysis of variance followed by Tukey’s post-tests. (PNG 231 kb)
Supplementary Fig. 2
HFD effects on duodenal motility. Pictures of duodenal segments from SCD-M (a), HFD-M (b), SCD-F (e) and HFD-F (f) mice during motility assays. Corresponding spatiotemporal maps (c-d, g-h) were generated in the GIMM program. Pictures and corresponding spatiotemporal maps show distinct contractions in SCD mice of both sexes (a, c, e, g). In contrast, HFD mice duodenums lack distinct uniform contractions (b, d, f, h) and in some cases, oral parts were severely distended and unable to contract (red asterisks) or relax properly. (PNG 1463 kb)
Supplementary Fig. 3
HFD determines microbial community composition. PCoA plot (a) shows gut microbial samples from both male and female mice fed SCD and HFD (PNG 305 kb)
Rights and permissions
About this article
Cite this article
Nyavor, Y., Estill, R., Edwards, H. et al. Intestinal nerve cell injury occurs prior to insulin resistance in female mice ingesting a high-fat diet. Cell Tissue Res 376, 325–340 (2019). https://doi.org/10.1007/s00441-019-03002-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00441-019-03002-0