Applied Microbiology and Biotechnology

, Volume 102, Issue 21, pp 9303–9316 | Cite as

Roxithromycin regulates intestinal microbiota and alters colonic epithelial gene expression

  • Cheng Zhang
  • Xuanwei Li
  • Liu Liu
  • Lijuan Gao
  • Shiyi Ou
  • Jianming LuoEmail author
  • Xichun PengEmail author
Genomics, transcriptomics, proteomics


The specialty of gastroenterology will be affected profoundly by the ability to modify the gastrointestinal microbiota through the use of antibiotics. This study investigated the in vivo effect of roxithromycin on gut bacteria and gene expression of colonic epithelial cells (CECs) using microbial 16S rDNA and colonic epithelial cell RNA sequencing, respectively. The results showed that roxithromycin distinctly lowered the microbial diversity in both the small intestine and cecum and altered the compositions of bacteria at both the phylum and genus levels, including the reduction of some bacteria beneficial to the hosts’ health. Eight decreased and 8 increased genera in the small intestine and 17 decreased and 4 increased genera of bacteria in the cecum were most affected by roxithromycin consumption. This consumption further altered the CECs’ expression of multiple genes. Thirty-one genes, which were significantly enriched in seven KEGG pathways and related to immune response, wound healing, and fibrosis, were significantly affected. Roxithromycin ingestion in healthy hosts, therefore, might lead to some undesirable consequences via affecting hosts’ gut microbiota and CECs. Our work offers a more comprehensive understanding of the impact of consuming roxithromycin on human health.


Roxithromycin Gut microbiota Colonic epithelial cells Gene expression profile 



We thank Ruixia Qiu and Bing Yu from the Department of Food Science and Engineering, Jinan University, for their contributions to the experiments of this study.


The program was funded by the National Natural Science Funds (No. 31471589) and the Fundamental Research Funds for the Central Universities (No. 21615404).

Compliance with ethical standard

Conflict of interest

All authors declare that they have no conflict of interest.

Ethical approval

All Institutional Animal Care and Use Committee of Jinan University guidelines for the care and use of animals were followed.

Supplementary material

253_2018_9257_MOESM1_ESM.xlsx (1 mb)
ESM 1 (XLSX 1058 kb)


  1. Adzemovic MZ, Öckinger J, Zeitelhofer M, Hochmeister S, Beyeen AD, Paulson A, Gillett A, Thessen Hedreul M, Covacu R, Lassmann H, Olsson T, Jagodic M (2012) Expression of Ccl11 associates with immune response modulation and protection against neuroinflammation in rats. PLoS One 7:e39794CrossRefGoogle Scholar
  2. Amato KR, Yeoman CJ, Kent A, Righini N, Carbonero F, Estrada A, Gaskins HR, Stumpf RM, Yildirim S, Torralba M, Gillis M, Wilson BA, Nelson KE, White BA, Leigh SR (2013) Habitat degradation impacts black howler monkey (Alouatta pigra) gastrointestinal microbiomes. ISME J 7:1344–1353CrossRefGoogle Scholar
  3. Avery D, Govindaraju P, Jacob M, Todd L, Monslow J, Puré E (2017) Extracellular matrix directs phenotypic heterogeneity of activated fibroblasts. Matrix Biol 67:90–106. CrossRefPubMedGoogle Scholar
  4. Badal S, Delgoda R (2014) Role of the modulation of CYP1A1 expression and activity in chemoprevention. J Appl Toxicol 34:743–753CrossRefGoogle Scholar
  5. Bauernfeind A (1993) In-vitro activity of dirithromycin in comparison with other new and established macrolides. J Antimicrob Chemoth 31:39–49CrossRefGoogle Scholar
  6. Benarafa C, Wolf M (2015) CXCL14: the Swiss army knife chemokine. Oncotarget 6:34065–34066CrossRefGoogle Scholar
  7. Botes M, van Reenen CA, Dicks LM (2008) Evaluation of Enterococcus mundtii ST4SA and Lactobacillus plantarum 423 as probiotics by using a gastro-intestinal model with infant milk formulations as substrate. Int J Food Microbiol 128:362–370CrossRefGoogle Scholar
  8. Brook I (2007) Anaerobic infections diagnosis and management. Informa Healthcare USA Inc., New YorkCrossRefGoogle Scholar
  9. Bryskier A (1998) Roxithromycin: review of its antimicrobial activity. J Antimicrob Chemoth 41:1–21CrossRefGoogle Scholar
  10. Bryskier A, Agouridas C, Gasc JC (1993) Classification of macrolide antibiotics. In: Bryskier AJ, Butzler JP, Neu HC, Tulkens PM (eds) Macrolides, chemistry, pharmacology and clinical uses. Arnette-Blackwell, Paris, pp 5–66Google Scholar
  11. Caporaso JG, Kuczynski J, Stombaugh J, Bittinger K, Bushman FD, Costello EK, Fierer N, Pena AG, Goodrich JK, Gordon JI, Huttley GA, Kelley ST, Knights D, Koenig JE, Ley RE, Lozupone CA, McDonald D, Muegge BD, Pirrung M, Reeder J, Sevinsky JR, Turnbaugh PJ, Walters WA, Widmann J, Yatsunenko T, Zaneveld J, Knight R (2010) QIIME allows analysis of high-throughput community sequencing data. Nat Methods 7:335–336CrossRefGoogle Scholar
  12. Chen T, Cheng G, Ahmed S, Wang Y, Wang X, Hao H, Yuan Z (2017) New methodologies in screening of antibiotic residues in animal-derived foods: biosensors. Talanta 175:435–442CrossRefGoogle Scholar
  13. Conly J, Johnston B (2005) Where are all the new antibiotics? The new antibiotic paradox. Can J Infect Dis Med Microbiol 16:159–160CrossRefGoogle Scholar
  14. Dally J, Khan JS, Voisey A, Charalambous C, John HL, Woods EL, Steadman R, Moseley R, Midgley AC (2017) Hepatocyte growth factor mediates enhanced wound healing responses and resistance to transforming growth factor-β1-driven myofibroblast differentiation in oral mucosal fibroblasts. Int J Mol Sci 18:E1843CrossRefGoogle Scholar
  15. Dang H, Wang S, Yang L, Fang F, Xu F (2012) Upregulation of Shh and Ptc1 in hyperoxia-induced acute lung injury in neonatal rats. Mol Med Rep 6:297–302PubMedGoogle Scholar
  16. Dubreuil L (1987) In vitro comparison of roxithromycin and erythromycin against 900 anaerobic bacterial strains. J Antimicrob Chemoth 20:13–19CrossRefGoogle Scholar
  17. Fan B, Ma L, Li Q, Wang L, Zhou J, Wu J (2013) Role of PDGFs/PDGFRs signaling pathway in myocardial fibrosis of DOCA/salt hypertensive rats. Int J Clin Exp Pathol 7:16–27PubMedPubMedCentralGoogle Scholar
  18. Guo H, Wang Y, Zhao Z, Shao X (2015) Platelet factor 4 limits Th17 differentiation and ischaemia-reperfusion injury after liver transplantation in mice. Scand J Immunol 81:129–134CrossRefGoogle Scholar
  19. Hecht DW, Osmolski JR (2003) Activities of garenoxacin (BMS-284756) and other agents against anaerobic clinical isolates. Antimicrob Agents Chemother 47:910–916CrossRefGoogle Scholar
  20. Hsu HS, Liu CC, Lin JH, Hsu TW, Hsu JW, Su K, Hung SC (2017) Involvement of ER stress, PI3K/AKT activation, and lung fibroblast proliferation in bleomycin-induced pulmonary fibrosis. Sci Rep 7:14272CrossRefGoogle Scholar
  21. Huang J, Lin X, Xue B, Luo J, Gao L, Wang Y, Peng X (2016) Impact of polyphenols combined with high-fat diet on rats’ gut microbiota. J Funct Foods 26:763–771CrossRefGoogle Scholar
  22. Jain R, Danziger LH (2004) The macrolide antibiotics: a pharmacokinetic and pharmacodynamic overview. Curr Pharm Des 10:3045–3053CrossRefGoogle Scholar
  23. Jiang XT, Peng X, Deng GH, Sheng HF, Wang Y, Zhou HW, Tam NFY (2013) Illumina sequencing of 16S rRNA tag revealed spatial variations of bacterial communities in a mangrove wetland. Microbial Ecol 66:219–225CrossRefGoogle Scholar
  24. Jung SP, Kang H (2014) Assessment of microbial diversity bias associated with soil heterogeneity and sequencing resolution in pyrosequencing analyses. J Microbiol 52:574–580CrossRefGoogle Scholar
  25. Kim DY, Takeuchi K, Ishinaga H, Kishioka C, Suzuki S, Basbaum C, Majima Y (2004) Roxithromycin suppresses mucin gene expression in epithelial cells. Pharmacol 72:6–11CrossRefGoogle Scholar
  26. Kim S, Covington A, Pamer EG (2017) The intestinal microbiota: antibiotics, colonization resistance, and enteric pathogens. Immunol Rev 279:90–105CrossRefGoogle Scholar
  27. Konno S, Adachi M, Asano K, Kawazoe T, Okamoto K, Takahashi T (1992) Influences of roxithromycin on cell-mediated immune responses. Life Sci 51:PL107–PL112CrossRefGoogle Scholar
  28. Langille MG, Zaneveld J, Caporaso JG, McDonald D, Knights D, Reyes JA, Clemente JC, Burkepile DE, Vega Thurber RL, Knight R, Beiko RG, Huttenhower C (2013) Predictive functional profiling of microbial communities using 16S rRNA marker gene sequences. Nat Biotechnol 31:814–821CrossRefGoogle Scholar
  29. Langmead B, Salzberg SL (2012) Fast gapped-read alignment with Bowtie 2. Nat Methods 9:357–359CrossRefGoogle Scholar
  30. Li B, Dewey CN (2011) RSEM: accurate transcript quantification from RNA-Seq data with or without a reference genome. BMC Bioinformatics 12:323CrossRefGoogle Scholar
  31. Marra F, DeFranco R, Grappone C, Milani S, Pinzani M, Pellegrini G, Laffi G, Gentilini P (1998) Expression of the thrombin receptor in human liver: up-regulation during acute and chronic injury. Hepatology 27:426–471CrossRefGoogle Scholar
  32. Migone TS, Zhang J, Luo X, Zhuang L, Chen C, Hu B, Hong JS, Perry JW, Chen SF, Zhou JX, Cho YH, Ullrich S, Kanakaraj P, Carrell J, Boyd E, Olsen HS, Hu G, Pukac L, Liu D, Ni J, Kim S, Gentz R, Feng P, Moore PA, Ruben SM, Wei P (2002) TL1A is a TNF-like ligand for DR3 and TR6/DcR3 and functions as a T cell costimulator. Immunity 16:479–492CrossRefGoogle Scholar
  33. Mohammadi H, Pinto VI, Wang Y, Hinz B, Janmey PA, McCulloch CA (2015) Filamin A mediates wound closure by promoting elastic deformation and maintenance of tension in the collagen matrix. J Invest Dermatol 135:2852–2861CrossRefGoogle Scholar
  34. Molloy EL, Adams A, Moore JB, Masterson JC, Madrigal-Estebas L, Mahon BP, O'Dea S (2008) BMP4 induces an epithelial-mesenchymal transition-like response in adult airway epithelial cells. Growth Factors 26:12–22CrossRefGoogle Scholar
  35. Montpas N, St-Onge G, Nama N, Rhainds D, Benredjem B, Girard M, Hickson G, Pons V, Heveker N (2017) Ligand-specific conformational transitions and intracellular transport required for atypical chemokine receptor 3-mediated chemokine scavenging. J Biol Chem 293:893–905. CrossRefPubMedPubMedCentralGoogle Scholar
  36. Muraoka H, Ogawa M, Miyazaki S, Tsuji A, Kaneko Y, Goto S (1988) Bacteriological evaluation of a new macrolide, RU 28965. Chemotherapy (Tokyo) 36:18–34Google Scholar
  37. Neary R, Watson CJ, Baugh JA (2015) Epigenetics and the overhealing wound: the role of DNA methylation in fibrosis. Fibrogenesis Tissue Repair 8:18CrossRefGoogle Scholar
  38. Nogacka AM, Salazar N, Arboleya S, Suárez M, Fernández N, Solís G, de Los Reyes-Gavilán CG, Gueimonde M (2017) Early microbiota, antibiotics and health. Cell Mol Life Sci 75:83–91. CrossRefPubMedGoogle Scholar
  39. Philp CJ, Siebeke I, Clements D (2017) ECM crosslinking enhances fibroblast growth and protects against matrix proteolysis in lung fibrosis. Am J Respir Cell Mol Biol 58:594–603. CrossRefGoogle Scholar
  40. Puri SK, Lassman HB (1987) Roxithromycin: a pharmacokinetic review of a macrolide. J Antimicrob Chemother 20:89–100CrossRefGoogle Scholar
  41. Robinson MD, McCarthy DJ, Smyth GK (2010) edgeR: a Bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics 26:139–140CrossRefGoogle Scholar
  42. Sabtu N, Enoch DA, Brown NM (2015) Antibiotic resistance: what, why, where, when and how? Br Med Bull 116:105–113PubMedGoogle Scholar
  43. Sato Y, Yanagita M (2017) Resident fibroblasts in the kidney: a major driver of fibrosis and inflammation. Inflamm Regen 37:17CrossRefGoogle Scholar
  44. Schülin T, Wennersten, CB, Eliopoulos GM, Moellering R (1996) In vitro activity of RU 64004 against Gram-positive bacteria. In: Abstracts of the 36th Interscience Conference on Antimicrobial Agents and Chemotherapy: 15–18 September 1996, Ernest N. Morial Convention Center, New Orleans, Louisiana. American Society for Microbiology, Washington DC, Abstract F-220, p 138Google Scholar
  45. Segata N, Izard J, Waldron L, Gevers D, Miropolsky L, Garrett WS, Huttenhower C (2011) Metagenomic biomarker discovery and explanation. Genome Biol 12:R60CrossRefGoogle Scholar
  46. Shehadeh LA, Webster KA, Hare JM, Vazquez-Padron RI (2011) Dynamic regulation of vascular myosin light chain (MYL9) with injury and aging. PLoS One 6:e25855CrossRefGoogle Scholar
  47. Tao Y, Yu G, Chen D, Pan Y, Liu Z, Wei H, Peng D, Huang L, Wang Y, Yuan Z (2012) Determination of 17 macrolide antibiotics and avermectins residues in meat with accelerated solvent extraction by liquid chromatography-tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci 897:64–71CrossRefGoogle Scholar
  48. Wang J, Leung D (2007) Analyses of macrolide antibiotic residues in eggs, raw milk, and honey using both ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry and high-performance liquid chromatography/tandem mass spectrometry. Rapid Commun Mass Spectrom 21:3213–3222CrossRefGoogle Scholar
  49. Wang Y, Wang GR, Shelby A, Shoemaker NB, Salyers AA (2003) A newly discovered Bacteroides conjugative transposon, CTnGERM1, contains genes also found in Gram-positive bacteria. Appl Environ Microbiol 69:4595–4603CrossRefGoogle Scholar
  50. Xie C, Mao X, Huang J, Ding Y, Wu J, Dong S, Kong L, Gao G, Li CY, Wei L (2011) KOBAS 2.0: a web server for annotation and identification of enriched pathways and diseases. Nucleic Acids Res 39:W316–W322CrossRefGoogle Scholar
  51. Yamamoto S, Nakajima K, Kohsaka S (2010) Macrophage-colony stimulating factor as an inducer of microglial proliferation in axotomized rat facial nucleus. J Neurochem 115:1057–1067CrossRefGoogle Scholar
  52. Yao T, Kojima Y, Koyanagi A, Yokoi H, Saito T, Kawano K, Furukawa M, Kusunoki T, Ikeda K (2009) Eotaxin-1, -2, and -3 immunoreactivity and protein concentration in the nasal polyps of eosinophilic chronic rhinosinusitis patients. Laryngoscope 119:1053–1059CrossRefGoogle Scholar
  53. Young RA, Gonzalez JP, Sorkin EM (1989) Roxithromycin. A review of its antibacterial activity, pharmacokinetic properties and clinical efficacy. Drugs 37:8–41CrossRefGoogle Scholar
  54. Zhang C, Li S, Yang L, Huang P, Li W, Wang S, Zhao G, Zhang M, Pang X, Yan Z, Liu Y, Zhao L (2013) Structural modulation of gut microbiota in life-long calorie-restricted mice. Nat Commun 4:2163CrossRefGoogle Scholar
  55. Zhou W, Ling Y, Liu T, Zhang Y, Li J, Li H, Wu W, Jiang S, Feng F, Yuan F, Zhang F (2017) Simultaneous determination of 16 macrolide antibiotics and 4 metabolites in milk by using Quick, Easy, Cheap, Effective, Rugged, and Safe extraction (QuEChERS) and high performance liquid chromatography tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci 1061-1062:411–420CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Food Science and EngineeringJinan UniversityGuangzhouChina
  2. 2.Department of Biomedical Engineering, College of Life Science and TechnologyJinan UniversityGuangzhouChina

Personalised recommendations