Quercetin improves and protects Calu-3 airway epithelial barrier function
K M Diguilio, E Rybakovsky, M C Valenzano, H H Nguyen, E A Del Rio, E Newberry, R Spadea, J M Mullin
Frontiers in Cell and Developmental Biology, doi:10.3389/fcell.2023.1271201
Introduction: In light of the impact of airway barrier leaks in COVID-19 and the significance of vitamin D in COVID-19 outcomes, including airway barrier protection, we investigated whether the very common dietary flavonoid quercetin could also be efficacious in supporting airway barrier function. Methods: To address this question, we utilized the widely used airway epithelial cell culture model, Calu-3.
Results: We observed that treating Calu-3 cell layers with quercetin increased transepithelial electrical resistance while simultaneously reducing transepithelial leaks of 14C-D-mannitol (Jm) and 14C-inulin. The effects of quercetin were concentration-dependent and exhibited a biphasic time course. These effects of quercetin occurred with changes in tight junctional protein composition as well as a partial inhibition of cell replication that resulted in decreased linear junctional density. Both of these effects potentially contribute to improved barrier function. Quercetin was equally effective in reducing the barrier compromise caused by the pro-inflammatory cytokine TNF-α, an action that seemed to derive, in part, from reducing the elevation of ERK 1/2 caused by TNF-α. Discussion: Quercetin improved Calu-3 barrier function and reduced TNF-α-induced barrier compromise, mediated in part by changes in the tight junctional complex.
Data availability statement The raw data supporting the conclusion of this article will be made available by the authors, without undue reservation.
Ethics statement Ethical approval was not required for the studies on humans in accordance with the local legislation and institutional requirements because only commercially available established cell lines were used.
Author contributions KD: writing-original draft, writing-review and editing, and investigation. ER: investigation and writing-review and editing. MV: investigation and writing-review and editing. HN: investigation and writing-review and editing. ED: investigation and writing-review and editing. EN: investigation and writing-review and editing. RS: investigation and writing-review and editing. JM: writing-review and editing, conceptualization, supervision, and writing-original draft.
Conflict of interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
Publisher's note All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors, and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.
Supplementary material The Supplementary Material for this article can be..
References
Al-Khayri, Sahana, Nagella, Joseph, Alessa et al., Flavonoids as potential anti-inflammatory molecules: a review, Molecules,
doi:10.3390/molecules27092901
Amasheh, Luettig, Amasheh, Zeitz, Fromm et al., Effects of quercetin studied in colonic HT-29/B6 cells and rat intestine in vitro, Ann. N. Y. Acad. Sci,
doi:10.1111/j.1749-6632.2012.06609x
Amasheh, Schlichter, Amasheh, Mankertz, Zeitz et al., Quercetin enhances epithelial barrier function and increases claudin-4 expression in Caco-2 cells, J. Nutr,
doi:10.1093/jn/138.6.1067
Amevor, Cui, Du, Ning, Deng et al., Supplementation of dietary quercetin and vitamin E promotes the intestinal structure and immune barrier integrity in aged breeder hens, Front. Immunol,
doi:10.3389/fimmu.2022.860889
Bae, Choe, Holick, Lim, Association of vitamin D status with COVID-19 and its severity: vitamin D and COVID-19: a narrative review, Rev. Endocr. Metab. Disord,
doi:10.1007/s11154-021-09705-6
Burton, A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid, Biochem. J,
doi:10.1042/bj0620315
Callaghan, Ferrick, Rybakovsky, Thomas, Mullin, Epithelial barrier function properties of the 16HBE14o-human bronchial epithelial cell culture model, Biosci. Rep,
doi:10.1042/BSR20201532
Carrasco-Pozo, Morales, Gotteland, Polyphenols protect the epithelial barrier function of Caco-2 cells exposed to indomethacin through the modulation of occludin and zonula occludens-1 expression, J. Agric. Food Chem,
doi:10.1021/jf400150p
Chuenkitiyanon, Pengsuparp, Jianmongkol, Protective effect of quercetin on hydrogen peroxide-induced tight junction disruption, Int. J. Toxicol,
doi:10.1177/1091581810366487
Crews, Alessandrini, Erikson, Erks: their fifteen minutes has arrived, Cell Growth Differ
Diguilio, Rybakovsky, Abdavies, Chamoun, Flounders et al., Micronutrient improvement of epithelial barrier function in various disease states: a case for adjuvant therapy, Int. J. Mol. Sci,
doi:10.3390/ijms23062995
Fan, Li, Zhang, Zhao, Wang, Pretreatment of IEC-6 cells with quercetin and myricetin resists the indomethacin-induced barrier dysfunction via attenuating the calcium-mediated JNK/Src activation, Food Chem. Toxicol,
doi:10.1016/j.fct.2020.111896
Fan, Zhang, Zhao, Zhang, The impact of heat treatment of quercetin and myricetin on their activities to alleviate the acrylamide-induced cytotoxicity and barrier loss in IEC-6 cells, Plant Foods Hum. Nutr,
doi:10.1007/s11130-022-00994-z
Gamero-Estevez, Andonian, Jean-Claude, Gupta, Ryan, Temporal effects of quercetin on tight junction barrier properties and claudin expression and localization in MDCK II cells, Int. J. Mol. Sci,
doi:10.3390/ijms20194889
Gao, Raduka, Rezaee, Vitamin D 3 protects against respiratory syncytial virus-induced barrier dysfunction in airway epithelial cells via PKA signaling pathway, Eur. J. Cell Biol,
doi:10.1016/j.ejcb.2023.151336
Kreuzfelder, Joka, Keinecke, Obertacke, Schmit-Neuerburg et al., Adult respiratory distress syndrome as a specific manifestation of a general permeability defect in trauma patients, Am. Rev. Respir. Dis,
doi:10.1164/ajrccm/137.1.95
Leyva-López, Gutierrez-Grijalva, Ambriz-Perez, Heredia, Flavonoids as cytokine modulators: a possible therapy for inflammation-related diseases, Int. J. Mol. Sci,
doi:10.3390/ijms17060921
Li, Mao, Gu, Wei, Gong, Quercetin protects retina external barrier from oxidative stress injury by promoting autophagy, Cutan. Ocul. Toxicol,
doi:10.1080/15569527.2020.1860082
Mercado, Valenzano, Jeffers, Sedlak, Cugliari et al., Enhancement of tight junctional barrier function by micronutrients: compound-specific effects on permeability and claudin composition, PLoS One,
doi:10.1371/journal.pone.0078775
Mullin, Ginanni, Laughlin, Protein kinase C activation increases transepithelial transport of biologically active insulin, Cancer Res
Mullin, Kampherstein, Laughlin, Clarkin, Miller et al., Overexpression of protein kinase C-delta increases tight junction permeability in LLC-PK1 epithelia, Am. J. Physiol,
doi:10.1152/ajpcell.1998.275.2.C544
Nakashima, Hisada, Goda, Tenno, Kotake et al., Opposing effect of naringenin and quercetin on the junctional compartment of MDCK II cells to modulate the tight junction, Nutrients,
doi:10.3390/nu12113285
Noureddine, Chalubinski, Wawrzyniak, The role of defective epithelial barriers in allergic lung disease and asthma development, J. Asthma Allergy,
doi:10.2147/JAA.S324080
Petecchia, Sabatini, Usai, Caci, Varesio et al., Cytokines induce tight junction disassembly in airway cells via an EGFR-dependent MAPK/ERK1/2-pathway, Lab. Invest,
doi:10.1038/labinvest.2012.67
Rasch, Schmidle, Sancak, Herner, Huberle et al., Increased extravascular lung water index (EVLWI) reflects rapid non-cardiogenic oedema and mortality in COVID-19 associated ARDS, Sci. Rep,
doi:10.1038/s41598-021-91043-3
Rastogi, Bhansali, Khare, Suri, Yaddanapudi et al., Short term, high-dose vitamin D supplementation for COVID-19 disease: a randomised, placebo-controlled, study (SHADE study), Postgrad. Med. J,
doi:10.1136/postgradmedj-2020-139065
Rybakovsky, Diguilio, Valenzano, Geagan, Pham et al., Calcitriol modifies tight junctions, improves barrier function, and reduces TNF-α-induced barrier leak in the human lung-derived epithelial cell culture model, 16HBE 14o, Physiol. Rep,
doi:10.14814/phy2.15592
Rybakovsky, Valenzano, Deis, Diguilio, Thomas et al., Improvement of human-oral-epithelial-barrier function and of tight junctions by micronutrients, J. Agric. Food Chem,
doi:10.1021/acs.jafc.7b04203
Sharma, Tripathi, Sharma, Dixit, Flavonoids modulate tight junction barrier functions in hyperglycemic human intestinal Caco-2 cells, Nutrition,
doi:10.1016/j.nut.2020.110792
Soler, Miller, Laughlin, Carp, Klurfeld et al., Increased tight junctional permeability is associated with the development of colon cancer, Carcinogenesis,
doi:10.1093/carcin/20.8.1425
Suzuki, Hara, 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,
doi:10.3945/jn.108.100867
Tripathi, Hazari, Mishra, Kumar, Sagi, Quercetin: a savior of alveolar barrier integrity under hypoxic microenvironment, Tissue Barriers,
doi:10.1080/21688370.2021.1883963
Valenzano, Diguilio, Mercado, Teter, To et al., Remodeling of tight junctions and enhancement of barrier integrity of the CACO-2 intestinal epithelial cell layer by micronutrients, PLoS One,
doi:10.1371/journal.pone.0133926
Virot, Mathien, Pointurier, Poidevin, Labro et al., Characterization of pulmonary impairment associated with COVID-19 in patients requiring mechanical ventilation, Rev. Bras. Ter. Intensiva,
doi:10.5935/0103-507X.20210007
Wan, Winton, Soeller, Tovey, Gruenert et al., Der p 1 facilitates transepithelial allergen delivery by disruption of tight junctions, J. Clin. Invest,
doi:10.1172/JCI5844
Xu, Qin, Xu, Yang, Chen et al., Dietary quercetin supplementation attenuates diarrhea, and intestinal damage by regulating gut microbiota in weanling piglets, Oxid. Med. Cell Longev,
doi:10.1155/2021/6221012
Zhang, Li, Liu, Wang, Quercetin effectively improves LPS-induced intestinal inflammation, pyroptosis, and disruption of the barrier function through the TLR4/NF-κB/NLRP3 signaling pathway in vivo and <i>in vitro</i>, Food Nutr. Res,
doi:10.29219/fnrv66.8948
Zhou, Xue, Jiang, Zhang, Wang et al., Beneficial effects of quercetin on microcystin-LR induced tight junction defects, Front. Pharmacol,
doi:10.3389/fphar.2021.733993