Top

International Journal of Colorectal Disease

Published in:

01-05-2017 | Original Article

Myrrh exerts barrier-stabilising and -protective effects in HT-29/B6 and Caco-2 intestinal epithelial cells

Authors: Rita Rosenthal, Julia Luettig, Nina A. Hering, Susanne M. Krug, Uwe Albrecht, Michael Fromm, Jörg-Dieter Schulzke

Published in: International Journal of Colorectal Disease | Issue 5/2017

Abstract

Purpose

Myrrh, the oleo-gum resin of Commiphora molmol, is well known for its anti-inflammatory properties. In different animal models, it protected against DSS-, TNBS- and oxazolone-induced colitis. To date, no information concerning the effect of myrrh on barrier properties are available. Thus, this study investigates the effect of myrrh on paracellular barrier function in the absence or presence of the pro-inflammatory cytokine TNFα.

Methods

Monolayers of human colon cell lines HT-29/B6 and Caco-2 were incubated with myrrh under control conditions or after challenge with the pro-inflammatory cytokine TNFα. Barrier function was analysed by electrophysiological and permeability measurements, Western blotting, immunostaining in combination with confocal microscopy, and freeze-fracture electron microscopy.

Results

In Caco-2 cells, myrrh induced an increase in transepithelial resistance (TER) which was associated with downregulation of the channel-forming tight junction (TJ) protein claudin-2 via inhibition of the PI3 kinase signalling pathway. In HT-29/B6 cells, myrrh had no effect on barrier properties under basic conditions, but protected against barrier damage induced by TNFα, as indicated by a decrease in TER and an increase in fluorescein permeability. The TNFα effect was associated with a redistribution of the sealing TJ protein claudin-1, an increase in the expression of claudin-2 and a change in TJ ultrastructure. Most importantly, all TNFα effects were inhibited by myrrh. The effect of myrrh on claudin-2 expression in this cell line was mediated via inhibition of the STAT6 pathway.

Conclusions

This study shows for the first time that myrrh exerts barrier-stabilising and TNFα-antagonising effects in human intestinal epithelial cell models via inhibition of PI3K and STAT6 signalling. This suggests therapeutic application of myrrh in intestinal diseases associated with barrier defects and inflammation.

Available only for authorised users

Turner JR (2009) Intestinal mucosal barrier function in health and disease. Nat Rev Immunol 9(11):799–809CrossRefPubMed

Furuse M et al (1993) Occludin: a novel integral membrane protein localizing at tight junctions. J Cell Biol 123:1777–1788CrossRefPubMed

Ikenouchi J et al (2005) Tricellulin constitutes a novel barrier at tricellular contacts of epithelial cells. J Cell Biol 171(6):939–945CrossRefPubMedPubMedCentral

Günzel D, Fromm M (2012) Claudins and other tight junction proteins. Compr Physiol 2(3):1819–1852PubMed

Martin-Padura I et al (1998) Junctional adhesion molecule, a novel member of the immunoglobulin superfamily that distributes at intercellular junctions and modulates monocyte transmigration. J Cell Biol 142(1):117–127CrossRefPubMedPubMedCentral

Zeissig S et al (2007) Changes in expression and distribution of claudin 2, 5 and 8 lead to discontinuous tight junctions and barrier dysfunction in active Crohn’s disease. Gut 56(1):61–72CrossRefPubMed

Heller F et al (2005) Interleukin-13 is the key effector Th2 cytokine in ulcerative colitis that affects epithelial tight junctions, apoptosis, and cell restitution. Gastroenterology 129(2):550–564CrossRefPubMed

Oshima T, Miwa H, Joh T (2008) Changes in the expression of claudins in active ulcerative colitis. J Gastroenterol Hepatol 23(Suppl 2):S146–S150CrossRefPubMed

Luettig J et al (2015) Claudin-2 as a mediator of leaky gut barrier during intestinal inflammation. Tissue Barriers 3(1–2):e977176CrossRefPubMedPubMedCentral

10.

Amasheh M et al (2008) Quercetin enhances epithelial barrier function and increases claudin-4 expression in Caco-2 cells. J Nutr 138(6):1067–1073PubMed

11.

Suzuki T, Hara H (2009) Quercetin enhances intestinal barrier function through the assembly of zonula [corrected] occludens-2, occludin, and claudin-1 and the expression of claudin-4 in Caco-2 cells. J Nutr 139(5):965–974CrossRefPubMed

12.

Amasheh M et al (2010) TNFalpha-induced and berberine-antagonized tight junction barrier impairment via tyrosine kinase, Akt and NFkappaB signaling. J Cell Sci 123:4145–4155CrossRefPubMed

13.

Luettig J, et al. (2016) The ginger component 6-shogaol prevents TNF-alpha-induced barrier loss via inhibition of PI3K/Akt and NF-kappaB signaling. Mol Nutr Food Res

14.

Shen T et al (2012) The genus Commiphora: a review of its traditional uses, phytochemistry and pharmacology. J Ethnopharmacol 142(2):319–330CrossRefPubMed

15.

Ford RA, Api AM, Letizia CS (1992) Monographs on fragrance raw materials. Food Chem Toxicol 30 Suppl:1S–138SPubMed

16.

Cheon JH et al (2006) Plant sterol guggulsterone inhibits nuclear factor-kappaB signaling in intestinal epithelial cells by blocking IkappaB kinase and ameliorates acute murine colitis. Inflamm Bowel Dis 12(12):1152–1161CrossRefPubMed

17.

Mencarelli A et al (2009) The plant sterol guggulsterone attenuates inflammation and immune dysfunction in murine models of inflammatory bowel disease. Biochem Pharmacol 78(9):1214–1223CrossRefPubMed

18.

Manjula N et al (2006) Inhibition of MAP kinases by crude extract and pure compound isolated from Commiphora mukul leads to down regulation of TNF-alpha, IL-1beta and IL-2. Int Immunopharmacol 6(2):122–132CrossRefPubMed

19.

Langhorst J et al (2013) Randomised clinical trial: a herbal preparation of myrrh, chamomile and coffee charcoal compared with mesalazine in maintaining remission in ulcerative colitis—a double-blind, double-dummy study. Aliment Pharmacol Ther 38(5):490–500CrossRefPubMed

20.

Langhorst J et al (2014) Distinct kinetics in the frequency of peripheral CD4+ T cells in patients with ulcerative colitis experiencing a flare during treatment with mesalazine or with a herbal preparation of myrrh, chamomile, and coffee charcoal. PLoS One 9(8):e104257CrossRefPubMedPubMedCentral

21.

Schmitz H et al (1999) Tumor necrosis factor-alpha (TNFalpha) regulates the epithelial barrier in the human intestinal cell line HT-29/B6. J Cell Sci 112:137–146PubMed

22.

Hidalgo IJ, Raub TJ, Borchardt RT (1989) Characterization of the human colon carcinoma cell line (Caco-2) as a model system for intestinal epithelial permeability. Gastroenterology 96(3):736–749CrossRefPubMed

23.

Günzel D et al (2009) Claudin-10 exists in six alternatively spliced isoforms that exhibit distinct localization and function. J Cell Sci 122(Pt 10):1507–1517CrossRefPubMed

24.

Rosenthal R et al (2010) Claudin-2, a component of the tight junction, forms a paracellular water channel. J Cell Sci 123(11):1913–1921CrossRefPubMed

25.

Rosen MJ et al (2011) STAT6 activation in ulcerative colitis: a new target for prevention of IL-13-induced colon epithelial cell dysfunction. Inflamm Bowel Dis 17(11):2224–2234CrossRefPubMedPubMedCentral

26.

Amasheh S et al (2002) Claudin-2 expression induces cation-selective channels in tight junctions of epithelial cells. J Cell Sci 115:4969–4976CrossRefPubMed

27.

Furuse M et al (2001) Conversion of zonulae occludentes from tight to leaky strand type by introducing claudin-2 into Madin-Darby canine kidney I cells. J Cell Biol 153(2):263–272CrossRefPubMedPubMedCentral

28.

Thongon N, Krishnamra N (2012) Apical acidity decreases inhibitory effect of omeprazole on Mg(2+) absorption and claudin-7 and -12 expression in Caco-2 monolayers. Exp Mol Med 44(11):684–693CrossRefPubMedPubMedCentral

29.

Kreusel KM et al (1991) Cl⁻ secretion in epithelial monolayers of mucus-forming human colon cells (HT-29/B6). Am J Phys 261(4):574–582

30.

Cui W et al (2010) Tumor necrosis factor alpha increases epithelial barrier permeability by disrupting tight junctions in Caco-2 cells. Braz J Med Biol Res 43(4):330–337CrossRefPubMed

31.

Gitter AH et al (2000) Epithelial barrier defects in HT-29/B6 colonic cell monolayers induced by tumor necrosis factor-alpha. Ann N Y Acad Sci 915:193–203CrossRefPubMed

32.

Mankertz J et al (2009) TNFalpha up-regulates claudin-2 expression in epithelial HT-29/B6 cells via phosphatidylinositol-3-kinase signaling. Cell Tissue Res 336(1):67–77CrossRefPubMed

33.

Epple HJ et al (2009) Impairment of the intestinal barrier is evident in untreated but absent in suppressively treated HIV-infected patients. Gut 58(2):220–227CrossRefPubMed

34.

Schumann M, et al. (2011) Cell polarity-determining proteins Par-3 and PP-1 are involved in epithelial tight junction defects in coeliac disease. Gut

35.

Prasad S et al (2005) Inflammatory processes have differential effects on claudins 2, 3 and 4 in colonic epithelial cells. Lab Investig 85(9):1139–1162CrossRefPubMed

36.

Suzuki T, Yoshinaga N, Tanabe S (2011) Interleukin-6 (IL-6) regulates claudin-2 expression and tight junction permeability in intestinal epithelium. J Biol Chem 286(36):31263–31271CrossRefPubMedPubMedCentral

37.

Rosen MJ et al (2013) STAT6 deficiency ameliorates severity of oxazolone colitis by decreasing expression of claudin-2 and Th2-inducing cytokines. J Immunol 190(4):1849–1858CrossRefPubMedPubMedCentral

38.

Dhawan P et al (2011) Claudin-2 expression increases tumorigenicity of colon cancer cells: role of epidermal growth factor receptor activation. Oncogene 30(29):3234–3247CrossRefPubMedPubMedCentral

39.

Escaffit F, Boudreau F, Beaulieu JF (2005) Differential expression of claudin-2 along the human intestine: implication of GATA-4 in the maintenance of claudin-2 in differentiating cells. J Cell Physiol 203(1):15–26CrossRefPubMed

40.

Holmes JL et al (2006) Claudin profiling in the mouse during postnatal intestinal development and along the gastrointestinal tract reveals complex expression patterns. Gene Expr Patterns 6(6):581–588CrossRefPubMed

41.

Mankertz J et al (2004) Functional crosstalk between Wnt signaling and Cdx-related transcriptional activation in the regulation of the claudin-2 promoter activity. Biochem Biophys Res Commun 314(4):1001–1007CrossRefPubMed

42.

Milatz S et al (2010) Claudin-3 acts as a sealing component of the tight junction for ions of either charge and uncharged solutes. Biochim Biophys Acta 1798(11):2048–2057CrossRefPubMed

43.

Piontek J et al (2011) Elucidating the principles of the molecular organization of heteropolymeric tight junction strands. Cell Mol Life Sci 68(23):3903–3918CrossRefPubMedPubMedCentral

44.

McCarthy KM et al (2000) Inducible expression of claudin-1-myc but not occludin-VSV-G results in aberrant tight junction strand formation in MDCK cells. J Cell Sci 113 Pt 19:3387–3398PubMed

45.

de Oliveira SS et al (2005) Claudins upregulation in human colorectal cancer. FEBS Lett 579(27):6179–6185CrossRefPubMed

46.

Schmitz H et al (2000) Epithelial barrier and transport function of the colon in ulcerative colitis. Ann N Y Acad Sci 915:312–326CrossRefPubMed

47.

Das P et al (2012) Comparative tight junction protein expressions in colonic Crohn’s disease, ulcerative colitis, and tuberculosis: a new perspective. Virchows Arch 460(3):261–270CrossRefPubMed

48.

Albrecht U et al (2014) Efficacy and safety of a herbal medicinal product containing myrrh, chamomile and coffee charcoal for the treatment of gastrointestinal disorders: a non-interventional study. BMJ Open Gastroenterol 1(1):e000015CrossRefPubMed

49.

Abdul-Ghani RA, Loutfy N, Hassan A (2009) Myrrh and trematodoses in Egypt: an overview of safety, efficacy and effectiveness profiles. Parasitol Int 58(3):210–214CrossRefPubMed

50.

Deng R (2007) Therapeutic effects of guggul and its constituent guggulsterone: cardiovascular benefits. Cardiovasc Drug Rev 25(4):375–390PubMed

Title: Myrrh exerts barrier-stabilising and -protective effects in HT-29/B6 and Caco-2 intestinal epithelial cells
Authors: Rita Rosenthal
Julia Luettig
Nina A. Hering
Susanne M. Krug
Uwe Albrecht
Michael Fromm
Jörg-Dieter Schulzke
Publication date: 01-05-2017
Publisher: Springer Berlin Heidelberg
Published in: International Journal of Colorectal Disease / Issue 5/2017
Print ISSN: 0179-1958
Electronic ISSN: 1432-1262
DOI: https://doi.org/10.1007/s00384-016-2736-x

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Myrrh exerts barrier-stabilising and -protective effects in HT-29/B6 and Caco-2 intestinal epithelial cells

Abstract

Purpose

Methods

Results

Conclusions

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 5/2017

The yet-to-be discovered mechanism of anastomotic leakage—comment to: Lack of evidence for tissue hypoxia as a contributing factor in anastomotic leak following colon anastomosis and segmental devascularization in rats

Comment on article entitled “The RIPASA score is sensitive and specific for the diagnosis of acute appendicitis in a western population”

The role of single-shot metronidazole in the prevention of Clostridium difficile infection following ileostomy reversal surgery

Risk factors and outcomes of acute lower gastrointestinal bleeding in intestinal Behçet’s disease

Rectal advancement flap for the treatment of complex cryptoglandular anal fistulas: a systematic review and meta-analysis

An 18-month study of the effects of IBD symptomatology and emotion regulation on depressed mood