Crocin alleviates lipopolysaccharide-induced acute respiratory distress syndrome by protecting against glycocalyx damage and suppressing inflammatory signaling pathways

  • Dong Zhang
  • Bo-yang Qi
  • Wei- wei Zhu
  • Xiao Huang
  • Xiao-zhi WangEmail author
Original Research Paper



To explore the mechanisms of crocin against glycocalyx damage and inflammatory injury in lipopolysaccharide (LPS)-induced acute respiratory distress syndrome (ARDS) mice and LPS-stimulated human umbilical vein endothelial cells (HUVECs).


Mice were randomly divided into control, LPS, and crocin + LPS (15, 30, and 60 mg/kg) groups. HUVECs were separated into eight groups: control, crocin, matrix metalloproteinase 9 inhibitor (MMP-9 inhib), cathepsin L inhibitor (CTL inhib), LPS, MMP-9 inhib + LPS, CTL inhib + LPS, and crocin + LPS. The potential cytotoxic effect of crocin on HUVECs was mainly evaluated through methylthiazolyldiphenyl-tetrazolium bromide assay. Histological changes were assessed via hemotoxylin and eosin staining. Lung capillary permeability was detected on the basis of wet–dry ratio and through fluorescein isothiocyanate-albumin assay. Then, protein levels were detected through Western blot analysis, immunohistochemical staining, and immunofluorescence.


This study showed that crocin can improve the pulmonary vascular permeability in mice with LPS-induced ARDS and inhibit the inflammatory signaling pathways of high mobility group box, nuclear factor κB, and mitogen-activated protein kinase in vivo and in vitro. Crocin also protected against the degradation of endothelial glycocalyx heparan sulfate and syndecan-4 by inhibiting the expressions of CTL, heparanase, and MMP-9 in vivo and in vitro. Overall, this study revealed the protective effects of crocin on LPS-induced ARDS and elaborated their underlying mechanism.


Crocin alleviated LPS-induced ARDS by protecting against glycocalyx damage and suppressing inflammatory signaling pathways.


Crocin Inflammatory responses Glycocalyx damage LPS ARDS 



This work was supported by funding from the National Natural Science Foundation of China (no.: 81670078).

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.


  1. 1.
    Lin X, Barravecchia M, Kothari P, Young JL, Dean DA. β1-Na(+), K(+)-ATPase gene therapy upregulates tight junctions to rescue lipopolysaccharide-induced acute lung injury. Gene Ther. 2016;23(6):489–99.PubMedCrossRefGoogle Scholar
  2. 2.
    Ma L, Zhao Y, Wang R, Chen T, Li W, Nan Y, et al. 3,5,4'-Tri-O-acetylresveratrol attenuates lipopolysaccharide-induced acute respiratory distress syndrome via MAPK/SIRT1 pathway. Mediators Inflamm. 2015;2015:143074.PubMedPubMedCentralGoogle Scholar
  3. 3.
    Yang Y, Haeger SM, Suflita MA, Zhang F, Dailey KL, Colbert JF, et al. Fibroblast growth factor signaling mediates pulmonary endothelial glycocalyx reconstitution. Am J Respir Cell Mol Biol. 2017;56(6):727–37.PubMedPubMedCentralCrossRefGoogle Scholar
  4. 4.
    Strand ME, Aronsen JM, Braathen B, Sjaastad I, Kvaløy H, Tønnessen T, et al. Shedding of syndecan-4 promotes immune cell recruitment and mitigates cardiac dysfunction after lipopolysaccharide challenge in mice. J Mol Cell Cardiol. 2015;88:133–44.PubMedCrossRefGoogle Scholar
  5. 5.
    McDonald KK, Cooper S, Danielzak L, Leask RL. Glycocalyx degradation induces a proinflammatory phenotype and increased leukocyte adhesion in cultured endothelial cells under flow. PLoS ONE. 2016;11(12):e0167576.PubMedPubMedCentralCrossRefGoogle Scholar
  6. 6.
    Marki A, Esko JD, Pries AR, Ley K. Role of the endothelial surface layer in neutrophil recruitment. J Leukoc Biol. 2015;98(4):503–15.PubMedPubMedCentralCrossRefGoogle Scholar
  7. 7.
    Iba T, Levy JH, Hirota T, Hiki M, Sato K, Murakami T, et al. Protection of the endothelial glycocalyx by antithrombin in an endotoxin-induced rat model of sepsis. Thromb Res. 2018;171:1–6.PubMedCrossRefGoogle Scholar
  8. 8.
    Yang Y, Schmidt EP. The endothelial glycocalyx: an important regulator of the pulmonary vascular barrier. Tissue Barriers. 2013;1(1):23494.PubMedCrossRefGoogle Scholar
  9. 9.
    Glasner DR, Ratnasiri K, Puerta-Guardo H, Espinosa DA, Beatty PR, Harris E. Dengue virus NS1 cytokine-independent vascular leak is dependent on endothelial glycocalyx components. PLoS Pathog. 2017;13(11):e1006673.PubMedPubMedCentralCrossRefGoogle Scholar
  10. 10.
    Reine TM, Lanzalaco F, Kristiansen O, Enget AR, Satchell S, Jenssen TG, et al. Matrix metalloproteinase-9 mediated shedding of syndecan-4 in glomerular endothelial cells. Microcirculation. 2019;31:e12534.CrossRefGoogle Scholar
  11. 11.
    Worthen GS, Schwab B, Elson EL, Downey GP. Mechanics of stimulated neutrophils: cell stiffening induces retention in capillaries. Science. 1989;245(4914):183–6.PubMedCrossRefGoogle Scholar
  12. 12.
    Feng Z, Wang JW, Wang Y, Dong WW, Xu ZF. Propofol protects lung endothelial barrier function by suppression of high-mobility group box 1 (HMGB1) release and mitochondrial oxidative damage catalyzed by HMGB1. Med Sci Monit. 2019;25:3199–211.PubMedPubMedCentralCrossRefGoogle Scholar
  13. 13.
    Pace E, Ferraro M, Di Vincenzo S, Siena L, Gjomarkaj M. Effects of ceftaroline on the innate immune and on the inflammatory responses of bronchial epithelial cells exposed to cigarette smoke. Toxicol Lett. 2016;258:216–26.PubMedCrossRefGoogle Scholar
  14. 14.
    Li L, Hu J, He T, Zhang Q, Yang X, Lan X, et al. P38/MAPK contributes to endothelial barrier dysfunction via MAP4 phosphorylation-dependent microtubule disassembly in inflammation-induced acute lung injury. Sci Rep. 2015;5:8895.PubMedPubMedCentralCrossRefGoogle Scholar
  15. 15.
    Xu GL, Qian ZY, Yu SQ, Gong ZN, Shen XC. Evidence of crocin against endothelial injury induced by hydrogen peroxide in vitro. J Asian Nat Prod Res. 2006;8(1–2):79–85.PubMedCrossRefGoogle Scholar
  16. 16.
    Lee IA, Lee JH, Baek NI, Kim DH. Antihyperlipidemic effect of crocin isolated from the fructus of gardenia jasminoides and its metabolite crocetin. Biol Pharm Bull. 2005;28(11):2106–10.PubMedCrossRefGoogle Scholar
  17. 17.
    Nie Z, Deng S, Zhang L, Chen S, Lu Q, Peng H. Crocin protects against dexamethasone-induced osteoblast apoptosis by inhibiting the ROS/Ca2+-mediated mitochondrial pathway. Mol Med Rep. 2019;20(1):401–8.PubMedPubMedCentralGoogle Scholar
  18. 18.
    Li S, Liu X, Lei J, Yang J, Tian P, Gao Y. Crocin protects podocytes against oxidative stress and inflammation induced by high glucose through inhibition of NF-κB. Cell Physiol Biochem. 2017;42(4):1481–92.PubMedCrossRefGoogle Scholar
  19. 19.
    Dianat M, Radan M, Badavi M, Mard SA, Bayati V, Ahmadizadeh M. Crocin attenuates cigarette smoke-induced lung injury and cardiac dysfunction by anti-oxidative effects: the role of Nrf2 antioxidant system in preventing oxidative stress. Respir Res. 2018;19(1):58.PubMedPubMedCentralCrossRefGoogle Scholar
  20. 20.
    Rajaei Z, Hadjzadeh MA, Nemati H, Hosseini M, Ahmadi M, Shafiee S. Antihyperglycemic and antioxidant activity of crocin in streptozotocin-induced diabetic rats. J Med Food. 2013;16(3):206–10.PubMedCrossRefGoogle Scholar
  21. 21.
    Wang L, Huang X, Kong G, Xu H, Li J, Hao D, et al. Ulinastatin attenuates pulmonary endothelial glycocalyx damage and inhibits endothelial heparanase activity in LPS-induced ARDS. Biochem Biophys Res Commun. 2016;478(2):669–75.PubMedCrossRefGoogle Scholar
  22. 22.
    Aeffner F, Bolon B, Davis IC. Mouse models of acute respiratory distress syndrome: a review of analytical approaches, pathologic features, and common measurements. Toxicol Pathol. 2015;43(8):1074–92.PubMedCrossRefGoogle Scholar
  23. 23.
    Li B, Lin Q, Hou Q, Yin C, Zhang L, Li Y. Alkannin attenuates lipopolysaccharide-induced lung injury in mice via Rho/ROCK/NF-κB pathway. J Biochem Mol Toxicol. 2019;29:e22323.Google Scholar
  24. 24.
    Mammoto A, Mammoto T, Kanapathipillai M, Wing Yung C, Jiang E, Jiang A, et al. Control of lung vascular permeability and endotoxin-induced pulmonary oedema by changes in extracellular matrix mechanics. Nat Commun. 2013;4:1759.PubMedCrossRefGoogle Scholar
  25. 25.
    Garsen M, Lenoir O, Rops AL, Dijkman HB, Willemsen B, van Kuppevelt TH, et al. Endothelin-1 induces proteinuria by heparanase-mediated disruption of the glomerular glycocalyx. J Am Soc Nephrol. 2016;27(12):3545–51.PubMedPubMedCentralCrossRefGoogle Scholar
  26. 26.
    Lambaerts K, Wilcox-Adelman SA, Zimmermann P. The signaling mechanisms of syndecan heparan sulfate proteoglycans. Curr Opin Cell Biol. 2009;21(5):662–9.PubMedPubMedCentralCrossRefGoogle Scholar
  27. 27.
    Ramani VC, Pruett PS, Thompson CA, DeLucas LD, Sanderson RD. Heparan sulfate chains of syndecan-1 regulate ectodomain shedding. J Biol Chem. 2012;287(13):9952–61.PubMedPubMedCentralCrossRefGoogle Scholar
  28. 28.
    Bortolotto V, Grilli M. Every Cloud has a silver lining: proneurogenic effects of Aβ oligomers and HMGB-1 via activation of the RAGE-NF-κB axis. CNS Neurol Disord Drug Targets. 2017;16(10):1066–79.PubMedCrossRefGoogle Scholar
  29. 29.
    Karuppagounder V, Arumugam S, Thandavarayan RA, Pitchaimani V, Sreedhar R, Afrin R, et al. Modulation of HMGB1 translocation and RAGE/NFκB cascade by quercetin treatment mitigates atopic dermatitis in NC/Nga transgenic mice. Exp Dermatol. 2015;24(6):418–23.PubMedCrossRefGoogle Scholar
  30. 30.
    Kolářová H, Ambrůzová B, Šindlerová L, Klinke A, Kubala L. Modulation of endothelial glycocalyx structure under inflammatory conditions. Mediators Inflamm. 2014;2014:694312.PubMedPubMedCentralCrossRefGoogle Scholar
  31. 31.
    Schmidt EP, Yang Y, Janssen WJ, Gandjeva A, Perez MJ, Barthel L, et al. The pulmonary endothelial glycocalyx regulates neutrophil adhesion and lung injury during experimental sepsis. Nat Med. 2012;18(8):1217–23.PubMedCrossRefGoogle Scholar
  32. 32.
    Chelazzi C, Villa G, Mancinelli P, De Gaudio AR, Adembri C. Glycocalyx and sepsis-induced alterations in vascular permeability. Crit Care. 2015;19:26.PubMedPubMedCentralCrossRefGoogle Scholar
  33. 33.
    Johansson PI, Stensballe J, Rasmussen LS, Ostrowski SR. A high admission syndecan-1 level, a marker of endothelial glycocalyx degradation, is associated with inflammation, protein C depletion, fibrinolysis, and increased mortality in trauma patients. Ann Surg. 2011;254(2):194–200.PubMedCrossRefGoogle Scholar
  34. 34.
    Santhosh MS, Sundaram MS, Sunitha K, Jnaneshwari S, Devaraja S, Kemparaju K, et al. Propensity of crocin to offset vipera russelli venom induced oxidative stress mediated neutrophil apoptosis: a biochemical insight. Cytotechnology. 2016;68(1):73–85.PubMedCrossRefGoogle Scholar
  35. 35.
    Ma Y, Yang X, Chatterjee V, Meegan JE, Beard RS Jr, Yuan SY. Role of neutrophil extracellular traps and vesicles in regulating vascular endothelial permeability. Front Immunol. 2019;10:1037.PubMedPubMedCentralCrossRefGoogle Scholar
  36. 36.
    Suzuki K, Okada H, Takemura G, Takada C, Kuroda A, Yano H, et al. Neutrophil elastase damages the pulmonary endothelial glycocalyx in lipopolysaccharide-induced experimental endotoxemia. Am J Pathol. 2019;189(8):1526–35.PubMedCrossRefGoogle Scholar
  37. 37.
    Chen L, Li W, Qi D, Wang D. Lycium barbarum polysaccharide protects against LPS-induced ARDS by inhibiting apoptosis, oxidative stress, and inflammation in pulmonary endothelial cells. Free Radic Res. 2018;52(4):480–90.PubMedCrossRefGoogle Scholar
  38. 38.
    Dull RO, Mecham I, McJames S. Heparan sulfates mediate pressure-induced increase in lung endothelial hydraulic conductivity via nitric oxide/reactive oxygen species. Am J Physiol Lung Cell Mol Physiol. 2007;292(6):L1452–L14581458.PubMedCrossRefGoogle Scholar
  39. 39.
    Kramer A, van den Hoven M, Rops A, Wijnhoven T, van den Heuvel L, Lensen J, et al. Induction of glomerular heparanase expression in rats with adriamycin nephropathy is regulated by reactive oxygen species and the renin-angiotensin system. J Am Soc Nephrol. 2006;17(9):2513–20.PubMedCrossRefGoogle Scholar
  40. 40.
    Kalantar M, Kalantari H, Goudarzi M, Khorsandi L, Bakhit S, Kalantar H. Crocin ameliorates methotrexate-induced liver injury via inhibition of oxidative stress and inflammation in rats. Pharmacol Rep. 2019;71(4):746–52.PubMedCrossRefGoogle Scholar
  41. 41.
    Song JW, Zullo JA, Liveris D, Dragovich M, Zhang XF, Goligorsky MS. Therapeutic restoration of endothelial glycocalyx in sepsis. J Pharmacol Exp Ther. 2017;361(1):115–21.PubMedPubMedCentralCrossRefGoogle Scholar

Copyright information

© Springer Nature Switzerland AG 2020

Authors and Affiliations

  • Dong Zhang
    • 1
  • Bo-yang Qi
    • 1
  • Wei- wei Zhu
    • 1
  • Xiao Huang
    • 1
  • Xiao-zhi Wang
    • 1
    Email author
  1. 1.Department of Respirator Medicine and Intensive Care UnitAffiliated Hospital of Binzhou Medical UniversityBinzhouChina

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