Hostname: page-component-76fb5796d-9pm4c Total loading time: 0 Render date: 2024-04-26T14:17:47.032Z Has data issue: false hasContentIssue false

An Unexpected Alteration Colonic Mucus Appearance in the Constipation Model via an Intestinal Microenvironment

Published online by Cambridge University Press:  30 May 2022

Han Gao
Affiliation:
Department of Physiology and Pathophysiology, School of Basic Medical Science, Capital Medical University, Beijing 100069, China
Chen-chen Gao
Affiliation:
Department of Physiology and Pathophysiology, School of Basic Medical Science, Capital Medical University, Beijing 100069, China
Tian-tian Wang
Affiliation:
Department of Physiology and Pathophysiology, School of Basic Medical Science, Capital Medical University, Beijing 100069, China
Lei Gao
Affiliation:
Department of Biomedical Informatics, School of Biomedical Engineering, Capital Medical University, Beijing 100069, China
Guang-wen Li
Affiliation:
Department of Physiology and Pathophysiology, School of Basic Medical Science, Capital Medical University, Beijing 100069, China
Liang-yun Jin
Affiliation:
Experimental Center for Morphological Research Platform, Capital Medical University, Beijing 100069, China
Cheng-wei He
Affiliation:
Department of Physiology and Pathophysiology, School of Basic Medical Science, Capital Medical University, Beijing 100069, China
Bo-ya Wang
Affiliation:
Undergraduate Student of 2018 Eight Program of Clinical Medicine, Peking University Health Science Center, Beijing 100069, China
Lucia Zhang
Affiliation:
Class of 2025, Loomis Chaffee School, 4 Batchelder Road, Windsor, CT 06095, USA
Yue-xin Guo
Affiliation:
Oral Medicine “5+3” process, School of Basic Medical Science, Capital Medical University, Beijing 100069, China
Rong-xuan Hua
Affiliation:
Department of Clinical Medicine, School of Basic Medical Science, Capital Medical University, Beijing 100069, China
Hong-wei Shang
Affiliation:
Experimental Center for Morphological Research Platform, Capital Medical University, Beijing 100069, China
Jing-dong Xu*
Affiliation:
Department of Physiology and Pathophysiology, School of Basic Medical Science, Capital Medical University, Beijing 100069, China
*
*Corresponding author: Jing-dong Xu, E-mail: xujingdong@163.com
Get access

Abstract

Due to the lack of research between the inner layers in the structure of colonic mucous and the metabolism of fatty acid in the constipation model, we aim to determine the changes in the mucous phenotype of the colonic glycocalyx and the microbial community structure following treatment with Rhubarb extract in our research. The constipation and treatment models are generated using adult male C57BL/6N mice. We perform light microscopy and transmission electron microscopy (TEM) to detect a Muc2-rich inner mucus layer attached to mice colon under different conditions. In addition, 16S rDNA sequencing is performed to examine the intestinal flora. According to TEM images, we demonstrate that Rhubarb can promote mucin secretion and find direct evidence of dendritic structure-linked mucus structures with its assembly into a lamellar network in a pore size distribution in the isolated colon section. Moreover, the diversity of intestinal flora has noticeable changes in constipated mice. The present study characterizes a dendritic structure and persistent cross-links have significant changes accompanied by the alteration of intestinal flora in feces in models of constipation and pretreatment with Rhubarb extract.

Type
Biological Applications
Copyright
Copyright © The Author(s), 2022. Published by Cambridge University Press on behalf of the Microscopy Society of America

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

Footnotes

These authors have contributed equally to this work and share the first authorship.

References

Allen, A, Cunliffe, WJ, Pearson, JP, Sellers, LA & Ward, R (1984). Studies on gastrointestinal mucus. Scand J Gastroenterol Suppl 93, 101113.Google ScholarPubMed
Ambort, D, Johansson, ME, Gustafsson, JK, Nilsson, HE, Ermund, A, Johansson, BR, et al. (2012). Calcium and pH-dependent packing and release of the gel-forming MUC2 mucin. Proc Natl Acad Sci U S A 109(15), 56455650. doi:10.1073/pnas.1120269109CrossRefGoogle ScholarPubMed
Anand, RJ, Kohler, JW, Cavallo, JA, Li, J, Dubowski, T & Hackam, DJ (2007). Toll-like receptor 4 plays a role in macrophage phagocytosis during peritoneal sepsis. J Pediatr Surg 42(6), 927932; discussion 933. doi:10.1016/j.jpedsurg.2007.01.023.CrossRefGoogle Scholar
Bachmann, O & Seidler, U (2011). News from the end of the gut–how the highly segmental pattern of colonic HCO(3)(−) transport relates to absorptive function and mucosal integrity. Biol Pharm Bull 34(6), 794802.CrossRefGoogle Scholar
Bajka, BH, Rigby, NM, Cross, KL, Macierzanka, A & Mackie, AR (2015). The influence of small intestinal mucus structure on particle transport ex vivo. Colloids Surf B Biointerfaces 135, 7380. doi:10.1016/j.colsurfb.2015.07.038CrossRefGoogle ScholarPubMed
Bernardazzi, C, Xu, H, Tong, H, Laubitz, D, Figliuolo da Paz, V, Curiel, L & Ghishan, FK (2020). An indisputable role of NHE8 in mucosal protection. Am J Physiol Gastrointest Liver Physiol 319(4), G421G431. doi:10.1152/ajpgi.00246.2020CrossRefGoogle ScholarPubMed
Bharucha, AE (2007). Constipation. Best Pract Res Clin Gastroenterol 21(4), 709731. doi:10.1016/j.bpg.2007.07.001CrossRefGoogle ScholarPubMed
Bharucha, AE, Pemberton, JH & Locke, GR 3rd. (2013). American gastroenterological association technical review on constipation. Gastroenterology 144(1), 218238. doi:10.1053/j.gastro.2012.10.028CrossRefGoogle ScholarPubMed
Birchenough, GM, Nystrom, EE, Johansson, ME & Hansson, GC (2016). A sentinel goblet cell guards the colonic crypt by triggering Nlrp6-dependent Muc2 secretion. Science 352(6293), 15351542. doi:10.1126/science.aaf7419CrossRefGoogle ScholarPubMed
Chassaing, B, Ley, RE & Gewirtz, AT (2014). Intestinal epithelial cell toll-like receptor 5 regulates the intestinal microbiota to prevent low-grade inflammation and metabolic syndrome in mice. Gastroenterology 147(6), 13631377.e1317. doi:10.1053/j.gastro.2014.08.033CrossRefGoogle ScholarPubMed
Chen, JQ, Chen, YY, Tao, HJ, Pu, ZJ, Shi, XQ, Zhang, J, et al. (2020). An integrated metabolomics strategy to reveal dose-effect relationship and therapeutic mechanisms of different efficacy of Rhubarb in constipation rats. J Pharm Biomed Anal 177, 112837. doi:10.1016/j.jpba.2019.112837CrossRefGoogle ScholarPubMed
Cullen, G & O'Donoghue, D (2007). Constipation and pregnancy. Best Pract Res Clin Gastroenterol 21(5), 807818. doi:10.1016/j.bpg.2007.05.005CrossRefGoogle ScholarPubMed
Deng, M, Scott, MJ, Loughran, P, Gibson, G, Sodhi, C, Watkins, S, et al. (2013). Lipopolysaccharide clearance, bacterial clearance, and systemic inflammatory responses are regulated by cell type-specific functions of TLR4 during sepsis. J Immunol 190(10), 51525160. doi:10.4049/jimmunol.1300496CrossRefGoogle ScholarPubMed
Dix, SR, Owen, HJ, Sun, R, Ahmad, A, Shastri, S, Spiewak, HL, et al. (2018). Structural insights into the function of type VI secretion system TssA subunits. Nat Commun 9(1), 4765. doi:10.1038/s41467-018-07247-1CrossRefGoogle ScholarPubMed
Ellingham, RB, Berry, M, Stevenson, D & Corfield, AP (1999). Secreted human conjunctival mucus contains MUC5AC glycoforms. Glycobiology 9(11), 11811189. doi:10.1093/glycob/9.11.1181CrossRefGoogle ScholarPubMed
Ermund, A, Schutte, A, Johansson, ME, Gustafsson, JK & Hansson, GC (2013). Studies of mucus in mouse stomach, small intestine, and colon. I. Gastrointestinal mucus layers have different properties depending on location as well as over the peyer's patches. Am J Physiol: Gastrointest Liver Physiol 305(5), G341G347. doi:10.1152/ajpgi.00046.2013Google Scholar
Estrada, JC, Brinn, NT & Bossen, EH (1985). A rapid method of staining ultrathin sections for surgical pathology TEM with the use of the microwave oven. Am J Clin Pathol 83(5), 639641.CrossRefGoogle ScholarPubMed
Fallon, RJ, Quirke, JF, Limper, J, Justus, C & Larkin, H (1991). The effects of buscopan compositum on calf nutritional diarrhoea. Vet Res Commun 15(6), 475482.Google ScholarPubMed
Gao, CC, Li, GW, Wang, TT, Gao, L, Wang, FF, Shang, HW, et al. (2021). Rhubarb extract relieves constipation by stimulating mucus production in the colon and altering the intestinal flora. Biomed Pharmacother 138, 111479. doi:10.1016/j.biopha.2021.111479CrossRefGoogle ScholarPubMed
Godl, K, Johansson, ME, Lidell, ME, Morgelin, M, Karlsson, H, Olson, FJ, et al. (2002). The N terminus of the MUC2 mucin forms trimers that are held together within a trypsin-resistant core fragment. J Biol Chem 277(49), 4724847256. doi:10.1074/jbc.M208483200CrossRefGoogle Scholar
Gong, XH, Li, Y, Zhang, RQ, Xie, XF, Peng, C & Li, YX (2015). The synergism mechanism of rhubarb anthraquinones on constipation elucidated by comparative pharmacokinetics of rhubarb extract between normal and diseased rats. Eur J Drug Metab Pharmacokinet 40(4), 379388. doi:10.1007/s13318-014-0216-7CrossRefGoogle ScholarPubMed
Grigor'eva, IN (2020). Gallstone disease, obesity and the Firmicutes/Bacteroidetes ratio as a possible biomarker of Gut dysbiosis. J Per Med 11(1). doi:10.3390/jpm11010013Google ScholarPubMed
Gustafsson, JK, Ermund, A, Ambort, D, Johansson, ME, Nilsson, HE, Thorell, K, et al. (2012 a). Bicarbonate and functional CFTR channel are required for proper mucin secretion and link cystic fibrosis with its mucus phenotype. J Exp Med 209(7), 12631272. doi:10.1084/jem.20120562Google ScholarPubMed
Gustafsson, JK, Ermund, A, Johansson, ME, Schutte, A, Hansson, GC & Sjovall, H (2012 b). An ex vivo method for studying mucus formation, properties, and thickness in human colonic biopsies and mouse small and large intestinal explants. Am J Physiol: Gastrointest Liver Physiol 302(4), G430G438. doi:10.1152/ajpgi.00405.2011Google Scholar
Hanson, B, Siddique, SM, Scarlett, Y & Sultan, S (2019). American gastroenterological association institute technical review on the medical management of opioid-induced constipation. Gastroenterology 156(1), 229253 e225. doi:10.1053/j.gastro.2018.08.018CrossRefGoogle ScholarPubMed
Hansson, GC & Johansson, ME (2010). The inner of the two Muc2 mucin-dependent mucus layers in colon is devoid of bacteria. Gut Microbes 1(1), 5154. doi:10.4161/gmic.1.1.10470CrossRefGoogle Scholar
Hong, Z, Chasan, B, Bansil, R, Turner, BS, Bhaskar, KR & Afdhal, NH (2005). Atomic force microscopy reveals aggregation of gastric mucin at low pH. Biomacromolecules 6(6), 34583466. doi:10.1021/bm0505843CrossRefGoogle ScholarPubMed
Hsu, CM, Yu, SC, Tsai, FJ & Tsai, Y (2013). Enhancement of rhubarb extract solubility and bioactivity by 2-hydroxypropyl-beta-cyclodextrin. Carbohydr Polym 98(2), 14221429. doi:10.1016/j.carbpol.2013.07.029CrossRefGoogle ScholarPubMed
Iwanaga, N, Nakamura, S, Oshima, K, Kajihara, T, Takazono, T, Miyazaki, T, et al. (2015). Macrolides promote CCL2-mediated macrophage recruitment and clearance of nasopharyngeal pneumococcal colonization in mice. J Infect Dis 212(7), 11501159. doi:10.1093/infdis/jiv157CrossRefGoogle ScholarPubMed
Jiang, D, Nelson, ML, Gally, F, Smith, S, Wu, Q, Minor, M, et al. (2012). Airway epithelial NF-kappaB activation promotes Mycoplasma pneumoniae clearance in mice. PLoS One 7(12), e52969. doi:10.1371/journal.pone.0052969.CrossRefGoogle ScholarPubMed
Jiang, F, Zhou, JY, Wu, J, Tian, F, Zhu, XX, Zhu, CL, et al. (2017). Yangyin runchang decoction improves intestinal motility in mice with atropine/diphenoxylate-induced slow-transit constipation. Evid Based Compl Altern Med 2017, 4249016. doi:10.1155/2017/4249016Google ScholarPubMed
Johansson, ME, Larsson, JM & Hansson, GC (2011). The two mucus layers of colon are organized by the MUC2 mucin, whereas the outer layer is a legislator of host-microbial interactions. Proc Natl Acad Sci U S A 108(Suppl 1), 46594665. doi:10.1073/pnas.1006451107CrossRefGoogle ScholarPubMed
Johansson, ME, Phillipson, M, Petersson, J, Velcich, A, Holm, L & Hansson, GC (2008). The inner of the two Muc2 mucin-dependent mucus layers in colon is devoid of bacteria. Proc Natl Acad Sci U S A 105(39), 1506415069. doi:10.1073/pnas.0803124105CrossRefGoogle Scholar
Johansson, ME, Sjovall, H & Hansson, GC (2013). The gastrointestinal mucus system in health and disease. Nat Rev Gastroenterol Hepatol 10(6), 352361. doi:10.1038/nrgastro.2013.35CrossRefGoogle ScholarPubMed
Johansson, ME, Thomsson, KA & Hansson, GC (2009). Proteomic analyses of the two mucus layers of the colon barrier reveal that their main component, the Muc2 mucin, is strongly bound to the fcgbp protein. J Proteome Res 8(7), 35493557. doi:10.1021/pr9002504CrossRefGoogle Scholar
Khalif, IL, Quigley, EM, Konovitch, EA & Maximova, ID (2005). Alterations in the colonic flora and intestinal permeability and evidence of immune activation in chronic constipation. Dig Liver Dis 37(11), 838849. doi:10.1016/j.dld.2005.06.008CrossRefGoogle ScholarPubMed
Kim, JE, Park, SH, Kwak, MH, Go, J, Koh, EK, Song, SH, et al. (2015). Characterization of changes in global genes expression in the distal colon of loperamide-induced constipation SD rats in response to the laxative effects of Liriope platyphylla. PLoS One 10(7), e0129664. doi:10.1371/journal.pone.0129664CrossRefGoogle Scholar
Liao, X & Wiedmann, TS (2006). Formation of cholesterol crystals at a mucin coated substrate. Pharm Res 23(10), 24132416. doi:10.1007/s11095-006-9004-7CrossRefGoogle Scholar
Linden, SK, Sutton, P, Karlsson, NG, Korolik, V & McGuckin, MA (2008). Mucins in the mucosal barrier to infection. Mucosal Immunol 1(3), 183197. doi:10.1038/mi.2008.5CrossRefGoogle ScholarPubMed
Lluch, J, Servant, F, Paisse, S, Valle, C, Valiere, S, Kuchly, C, et al. (2015). The characterization of novel tissue Microbiota using an optimized 16S metagenomic sequencing pipeline. PLoS One 10(11), e0142334. doi:10.1371/journal.pone.0142334Google ScholarPubMed
Lu, Y, Chen, J, Zheng, J, Hu, G, Wang, J, Huang, C, et al. (2016). Mucosal adherent bacterial dysbiosis in patients with colorectal adenomas. Sci Rep 6, 26337. doi:10.1038/srep26337CrossRefGoogle ScholarPubMed
Majewski, M, Sarosiek, I, Wallner, G, Edlavitch, SA & Sarosiek, J (2014). Stimulation of mucin, mucus, and viscosity during lubiprostone in patients with chronic constipation may potentially lead to increase of lubrication. Clin Transl Gastroenterol 5, e66. doi:10.1038/ctg.2014.19CrossRefGoogle ScholarPubMed
Muller-Lissner, S (2007). The difficult patient with constipation. Best Pract Res Clin Gastroenterol 21(3), 473484. doi:10.1016/j.bpg.2007.01.008CrossRefGoogle ScholarPubMed
Nishida, A, Lau, CW, Zhang, M, Andoh, A, Shi, HN, Mizoguchi, E & Mizoguchi, A (2012). The membrane-bound mucin Muc1 regulates T helper 17-cell responses and colitis in mice. Gastroenterology 142(4), 865874.e862. doi:10.1053/j.gastro.2011.12.036CrossRefGoogle ScholarPubMed
Pelaseyed, T, Bergstrom, JH, Gustafsson, JK, Ermund, A, Birchenough, GM, Schutte, A, et al. (2014). The mucus and mucins of the goblet cells and enterocytes provide the first defense line of the gastrointestinal tract and interact with the immune system. Immunol Rev 260(1), 820. doi:10.1111/imr.12182CrossRefGoogle ScholarPubMed
Qiao, H, Zhang, L, Shi, H, Song, Y & Bian, C (2018). Astragalus affects fecal microbial composition of young hens as determined by 16S rRNA sequencing. AMB Express 8(1), 70. doi:10.1186/s13568-018-0600-9CrossRefGoogle ScholarPubMed
Round, AN, Berry, M, McMaster, TJ, Corfield, AP & Miles, MJ (2004). Glycopolymer charge density determines conformation in human ocular mucin gene products: An atomic force microscope study. J Struct Biol 145(3), 246253. doi:10.1016/j.jsb.2003.10.029CrossRefGoogle Scholar
Round, AN, Berry, M, McMaster, TJ, Stoll, S, Gowers, D, Corfield, AP & Miles, MJ (2002). Heterogeneity and persistence length in human ocular mucins. Biophys J 83(3), 16611670. doi:10.1016/S0006-3495(02)73934-9CrossRefGoogle ScholarPubMed
Round, AN, McMaster, TJ, Miles, MJ, Corfield, AP & Berry, M (2007). The isolated MUC5AC gene product from human ocular mucin displays intramolecular conformational heterogeneity. Glycobiology 17(6), 578585. doi:10.1093/glycob/cwm027Google ScholarPubMed
Round, AN, Rigby, NM, Garcia de la Torre, A, Macierzanka, A, Mills, EN & Mackie, AR (2012). Lamellar structures of MUC2-rich mucin: A potential role in governing the barrier and lubricating functions of intestinal mucus. Biomacromolecules 13(10), 32533261. doi:10.1021/bm301024xCrossRefGoogle ScholarPubMed
Sellers, LA, Allen, A, Morris, ER & Ross-Murphy, SB (1991). The rheology of pig small intestinal and colonic mucus: Weakening of gel structure by non-mucin components. Biochim Biophys Acta 1115(2), 174179.CrossRefGoogle ScholarPubMed
Seva-Pereira, A, de Moraes, GR, de Oliveira, SP & Reyes, FG (1991). [Use of fiber enriched biscuit in the treatment of chronic intestinal constipation]. Rev Med 109(6), 265268.Google Scholar
Thornton, DJ & Sheehan, JK (2004). From mucins to mucus: Toward a more coherent understanding of this essential barrier. Proc Am Thorac Soc 1(1), 5461. doi:10.1513/pats.2306016CrossRefGoogle Scholar
van der Post, S, Thomsson, KA & Hansson, GC (2014). Multiple enzyme approach for the characterization of glycan modifications on the C-terminus of the intestinal MUC2mucin. J Proteome Res 13(12), 60136023. doi:10.1021/pr500874fCrossRefGoogle ScholarPubMed
Wang, X & Yin, J (2015). Complementary and alternative therapies for chronic constipation. Evid Based Complement Altern Med 2015, 396396. doi:10.1155/2015/396396.Google ScholarPubMed
Wei, L, Luo, Y, Zhang, X, Liu, Y, Gasser, M, Tang, F, et al. (2020). Topical therapy with rhubarb navel plasters in patients with chronic constipation: Results from a prospective randomized multicenter study. J Ethnopharmacol, 113096. doi:10.1016/j.jep.2020.113096Google ScholarPubMed
Wu, D, Xue, X, Gao, C, Liu, Y, Wang, T, Li, L, et al. (2019). Rhubarb-Evoke mucus secretion through aggregation and degranulation of mast cell in the colon of Rat: In vivo and ex vivo studies. Sci Rep 9(1), 19375. doi:10.1038/s41598-019-55937-7CrossRefGoogle ScholarPubMed
Xiao, B, Niu, X, Han, N, Wang, B, Du, P, Na, R, et al. (2016). Predictive value of the composition of the vaginal microbiota in bacterial vaginosis, a dynamic study to identify recurrence-related flora. Sci Rep 6, 26674. doi:10.1038/srep26674CrossRefGoogle ScholarPubMed
Xu, JD, Liu, S, Wang, W, Li, LS, Li, XF, Li, Y, et al. (2012). Emodin induces chloride secretion in rat distal colon through activation of mast cells and enteric neurons. Br J Pharmacol 165(1), 197207. doi:10.1111/j.1476-5381.2011.01573.xCrossRefGoogle ScholarPubMed
Xu, JD, Wang, W, Li, LS, Chen, X & Zhu, JX (2007). Involvement of endogenous prostaglandin in emodin-evoked rat colonic anion secretion. Biol Pharm Bull 30(11), 20582062. doi:10.1248/bpb.30.2058CrossRefGoogle ScholarPubMed
Zhai, X, Lin, D, Zhao, Y & Yang, X (2018). Bacterial cellulose relieves diphenoxylate-induced constipation in rats. J Agric Food Chem 66(16), 41064117. doi:10.1021/acs.jafc.8b00385CrossRefGoogle ScholarPubMed
Zhao, YL, Wang, JB, Zhou, GD, Shan, LM & Xiao, XH (2009). Investigations of free anthraquinones from rhubarb against alpha-naphthylisothiocyanate-induced cholestatic liver injury in rats. Basic Clin Pharmacol Toxicol 104(6), 463469. doi:10.1111/j.1742-7843.2009.00389.xCrossRefGoogle ScholarPubMed
Zhu, L, Liu, W, Alkhouri, R, Baker, RD, Bard, JE, Quigley, EM & Baker, SS (2014). Structural changes in the gut microbiome of constipated patients. Physiol Genomics 46(18), 679686. doi:10.1152/physiolgenomics.00082.2014.CrossRefGoogle ScholarPubMed