Abstract
Disruption of the endothelial barrier function and reduction in cell migration leads to endothelial dysfunction. One of the most abundant human milk oligosaccharides, 6′-sialylactose (6′-SL), is reported to exert various biological functions related to inflammatory responses. In this study, we evaluated the effects of 6′-SL on lipopolysaccharide (LPS)-induced inflammation caused by endothelial barrier damage. Our results showed that LPS at 500 ng/mL strongly not only abolished cell migration but also hyperactivated MAPK and NF-κB pathways. 6′-SL suppressed LPS-induced endothelial inflammation via ERK1/2, p38, and JNK MAPK pathways. 6′-SL supported endothelial junctions by upregulating PECAM-1 expression and mRNA levels of tight junctions, such as ZO-1 and occludin, which were downregulated by LPS stimulation. It significantly inhibited the nuclear translocation of NF-κB, along with the downregulation of inflammatory cytokines, including TNF-α, IL-1β, MCP-1, VCAM-1, and ICAM-1. Furthermore, 6′-SL abolished NF-κB-mediated STAT3 in controlling endothelial migration and hyperpermeability via downregulating STAT3 activation and nuclear translocation. Finally, LPS induced over-expression of VCAM-1 and ZO-1 disassembly in both atheroprone and atheroprotective areas of mouse aorta, which were reversed by 6′-SL treatment. Altogether, our findings suggest that 6′-SL is a potent therapeutic agent for modulating inflammatory responses and endothelial hyperpermeability.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Almalki SG, Agrawal DK (2017) ERK signaling is required for VEGF-A/VEGFR2-induced differentiation of porcine adipose-derived mesenchymal stem cells into endothelial cells. Stem Cell Res Ther 8:113. https://doi.org/10.1186/s13287-017-0568-4
Baek N, Sim S, Heo K-S (2018) LPS-stimulated macrophage activation affects endothelial dysfunction. JBV 48:23–30. https://doi.org/10.4167/jbv.2018.48.1.23
Birukova AA, Wu T, Tian Y, Meliton A, Sarich N, Tian X, Leff A, Birukov KG (2013) Iloprost improves endothelial barrier function in lipopolysaccharide-induced lung injury. Eur Respir J 41:165–176. https://doi.org/10.1183/09031936.00148311
Bode L, Kunz C, Muhly-Reinholz M, Mayer K, Seeger W, Rudloff S (2004) Inhibition of monocyte, lymphocyte, and neutrophil adhesion to endothelial cells by human milk oligosaccharides. Thromb Haemost 92:1402–1410. https://doi.org/10.1160/th04-01-0055
Bonetti PO, Lerman LO, Lerman A (2003) Endothelial dysfunction: a marker of atherosclerotic risk. Arterioscler Thromb Vasc Biol 23:168–175. https://doi.org/10.1161/01.atv.0000051384.43104.fc
Cerutti C, Ridley AJ (2017) Endothelial cell-cell adhesion and signaling. Exp Cell Res 358:31–38. https://doi.org/10.1016/j.yexcr.2017.06.003
Chang Z, Wang Y, Zhou X, Long JE (2018) STAT3 roles in viral infection: antiviral or proviral? Future Virol 13:557–574. https://doi.org/10.2217/fvl-2018-0033
Chen Y, He SD, Li XD, Hu ZL, Zhang C, Xu F (2020) Long noncoding RNA atlas of the inflammation caused by asthma in mice. Arch Pharm Res 43:421–432. https://doi.org/10.1007/s12272-020-01223-4
Chung TW, Kim EY, Kim SJ, Choi HJ, Jang SB, Kim KJ, Ha SH, Abekura F, Kwak CH, Kim CH, Ha KT (2017) Sialyllactose suppresses angiogenesis by inhibiting VEGFR-2 activation, and tumor progression. Oncotarget 8:58152–58162. https://doi.org/10.18632/oncotarget.16192
Dauphinee SM, Karsan A (2006) Lipopolysaccharide signaling in endothelial cells. Lab Invest 86:9–22. https://doi.org/10.1038/labinvest.3700366
Elbashir SM, Martinez J, Patkaniowska A, Lendeckel W, Tuschl T (2001) Functional anatomy of siRNAs for mediating efficient RNAi in Drosophila melanogaster embryo lysate. Embo J 20:6877–6888. https://doi.org/10.1093/emboj/20.23.6877
Heo KS, Lee H, Nigro P, Thomas T, Le NT, Chang E, Mcclain C, Reinhart-King CA, King MR, Berk BC, Fujiwara K, Woo CH, Abe J (2011) PKCζ mediates disturbed flow-induced endothelial apoptosis via p53 SUMOylation. J Cell Biol 193:867–884. https://doi.org/10.1083/jcb.201010051
Heo KS, Le NT, Cushman HJ, Giancursio CJ, Chang E, Woo CH, Sullivan MA, Taunton J, Yeh ET, Fujiwara K, Abe J (2015) Disturbed flow-activated p90RSK kinase accelerates atherosclerosis by inhibiting SENP2 function. J Clin Invest 125:1299–1310. https://doi.org/10.1172/jci76453
Huynh DTN, Heo KS (2019) Therapeutic targets for endothelial dysfunction in vascular diseases. Arch Pharm Res 42:848–861. https://doi.org/10.1007/s12272-019-01180-7
Huynh DTN, Heo KS (2021) Role of mitochondrial dynamics and mitophagy of vascular smooth muscle cell proliferation and migration in progression of atherosclerosis. Arch Pharm Res 44:1051–1061. https://doi.org/10.1007/s12272-021-01360-4
Huynh DTN, Heo K-S (2022) Therapeutic effects of ginsenosides on vascular smooth muscle cell phenotypic switching in vascular diseases. Cardiometab Syndr J 2:96–107
Huynh DTN, Jin Y, Myung CS, Heo KS (2020) Inhibition of p90RSK is critical to abolish Angiotensin II-induced rat aortic smooth muscle cell proliferation and migration. Biochem Biophys Res Commun 523:267–273. https://doi.org/10.1016/j.bbrc.2019.12.053
Jang C, Kim J, Kwon Y, Jo SA (2020) Telmisartan inhibits TNFα-induced leukocyte adhesion by blocking ICAM-1 expression in astroglial cells but not in endothelial cells. Biomol Ther (seoul) 28:423–430. https://doi.org/10.4062/biomolther.2020.119
Jeon H, Jin Y, Myung CS, Heo KS (2021) Ginsenoside-Rg2 exerts anti-cancer effects through ROS-mediated AMPK activation associated mitochondrial damage and oxidation in MCF-7 cells. Arch Pharm Res 44:702–712. https://doi.org/10.1007/s12272-021-01345-3
Jeong JH, Ojha U, Lee YM (2021) Pathological angiogenesis and inflammation in tissues. Arch Pharm Res 44:1–15. https://doi.org/10.1007/s12272-020-01287-2
Ji SY, Cha HJ, Molagoda IMN, Kim MY, Kim SY, Hwangbo H, Lee H, Kim GY, Kim DH, Hyun JW, Kim HS, Kim S, Jin CY, Choi YH (2021) Suppression of lipopolysaccharide-induced inflammatory and oxidative response by 5-aminolevulinic acid in RAW 264.7 macrophages and Zebrafish Larvae. Biomol Ther (seoul) 29:685–696. https://doi.org/10.4062/biomolther.2021.030
Jin BR, Kim HJ, Kim EY, Chung TW, Ha KT, An HJ (2019) 6’-Sialyllactose ameliorates in vivo and in vitro benign prostatic hyperplasia by regulating the E2F1/pRb-AR pathway. Nutrients. https://doi.org/10.3390/nu11092203
Jin Y, Huynh DTN, Myung CS, Heo KS (2021) Ginsenoside Rh1 prevents migration and invasion through mitochondrial ROS-mediated inhibition of STAT3/NF-κB signaling in MDA-MB-231 cells. Int J Mol Sci. https://doi.org/10.3390/ijms221910458
Jin Y, Huynh DTN, Heo KS (2022a) Ginsenoside Rh1 inhibits tumor growth in MDA-MB-231 breast cancer cells via mitochondrial ROS and ER stress-mediated signaling pathway. Arch Pharm Res 45:174–184. https://doi.org/10.1007/s12272-022-01377-3
Jin Y, Nguyen TLL, Myung CS, Heo KS (2022b) Ginsenoside Rh1 protects human endothelial cells against lipopolysaccharide-induced inflammatory injury through inhibiting TLR2/4-mediated STAT3, NF-kappaB, and ER stress signaling pathways. Life Sci 309:120973. https://doi.org/10.1016/j.lfs.2022.120973
Komarova YA, Kruse K, Mehta D, Malik AB (2017) Protein interactions at endothelial junctions and signaling mechanisms regulating endothelial permeability. Circ Res 120:179–206. https://doi.org/10.1161/circresaha.116.306534
Liu F, Shi K, Dong J, Jin Z, Wu Y, Cai Y, Lin T, Cai Q, Liu L, Zhang Y (2020) Ganoderic acid A attenuates high-fat-diet-induced liver injury in rats by regulating the lipid oxidation and liver inflammation. Arch Pharm Res 43:744–754. https://doi.org/10.1007/s12272-020-01256-9
Maas M, Stapleton M, Bergom C, Mattson DL, Newman DK, Newman PJ (2005) Endothelial cell PECAM-1 confers protection against endotoxic shock. Am J Physiol Heart Circ Physiol 288:H159-164. https://doi.org/10.1152/ajpheart.00500.2004
Naito Y, Ui-Tei K (2012) siRNA design software for a target gene-specific RNA interference. Front Genet 3:102. https://doi.org/10.3389/fgene.2012.00102
Nguyen TLL, Huynh DTN, Jin Y, Jeon H, Heo KS (2021) Protective effects of ginsenoside-Rg2 and -Rh1 on liver function through inhibiting TAK1 and STAT3-mediated inflammatory activity and Nrf2/ARE-mediated antioxidant signaling pathway. Arch Pharm Res 44:241–252. https://doi.org/10.1007/s12272-020-01304-4
Nguyen TLL, Jin Y, Kim L, Heo KS (2022) Inhibitory effects of 6’-sialyllactose on angiotensin II-induced proliferation, migration, and osteogenic switching in vascular smooth muscle cells. Arch Pharm Res 45:658–670. https://doi.org/10.1007/s12272-022-01404-3
Ni J, Lin M, Jin Y, Li J, Guo Y, Zhou J, Hong G, Zhao G, Lu Z (2019) Gas6 attenuates sepsis-induced tight junction injury and vascular endothelial hyperpermeability via the Axl/NF-κB signaling pathway. Front Pharmacol 10:662. https://doi.org/10.3389/fphar.2019.00662
Phipps KR, Baldwin NJ, Lynch B, Stannard DR, Šoltésová A, Gilby B, Mikš MH, Röhrig CH (2019) Toxicological safety evaluation of the human-identical milk oligosaccharide 6’-sialyllactose sodium salt. J Appl Toxicol 39:1444–1461. https://doi.org/10.1002/jat.3830
Privratsky JR, Newman PJ (2014) PECAM-1: regulator of endothelial junctional integrity. Cell Tissue Res 355:607–619. https://doi.org/10.1007/s00441-013-1779-3
Ramalingam P, Poulos MG, Lazzari E, Gutkin MC, Lopez D, Kloss CC, Crowley MJ, Katsnelson L, Freire AG, Greenblatt MB, Park CY, Butler JM (2020) Chronic activation of endothelial MAPK disrupts hematopoiesis via NFKB dependent inflammatory stress reversible by SCGF. Nat Commun 11:666. https://doi.org/10.1038/s41467-020-14478-8
Sodhi CP, Wipf P, Yamaguchi Y, Fulton WB, Kovler M, Niño DF, Zhou Q, Banfield E, Werts AD, Ladd MR, Buck RH, Goehring KC, Prindle T Jr, Wang S, Jia H, Lu P, Hackam DJ (2021) The human milk oligosaccharides 2’-fucosyllactose and 6’-sialyllactose protect against the development of necrotizing enterocolitis by inhibiting toll-like receptor 4 signaling. Pediatr Res 89:91–101. https://doi.org/10.1038/s41390-020-0852-3
Ten Bruggencate SJ, Bovee-Oudenhoven IM, Feitsma AL, Van Hoffen E, Schoterman MH (2014) Functional role and mechanisms of sialyllactose and other sialylated milk oligosaccharides. Nutr Rev 72:377–389. https://doi.org/10.1111/nure.12106
Van Nieuw Amerongen GP, Van Hinsbergh VW (2002) Targets for pharmacological intervention of endothelial hyperpermeability and barrier function. Vascul Pharmacol 39:257–272. https://doi.org/10.1016/s1537-1891(03)00014-4
Wang L, Astone M, Alam SK, Zhu Z, Pei W, Frank DA, Burgess SM, Hoeppner LH (2021) Suppressing STAT3 activity protects the endothelial barrier from VEGF-mediated vascular permeability. Dis Model Mech. https://doi.org/10.1242/dmm.049029
Wiciński M, Sawicka E, Gębalski J, Kubiak K, Malinowski B (2020) Human milk oligosaccharides: health benefits, potential applications in infant formulas, and pharmacology. Nutrients. https://doi.org/10.3390/nu12010266
Wong E, Xu F, Joffre J, Nguyen N, Wilhelmsen K, Hellman J (2021) ERK1/2 has divergent roles in LPS-induced microvascular endothelial cell cytokine production and permeability. Shock 55:349–356. https://doi.org/10.1097/shk.0000000000001639
Xu F, Zhou F (2020) Inhibition of microRNA-92a ameliorates lipopolysaccharide-induced endothelial barrier dysfunction by targeting ITGA5 through the PI3K/Akt signaling pathway in human pulmonary microvascular endothelial cells. Int Immunopharmacol 78:106060. https://doi.org/10.1016/j.intimp.2019.106060
Xu S, Pan X, Mao L, Pan H, Xu W, Hu Y, Yu X, Chen Z, Qian S, Ye Y, Huang Y, Pan J (2020) Phospho-Tyr705 of STAT3 is a therapeutic target for sepsis through regulating inflammation and coagulation. Cell Commun Signal 18:104. https://doi.org/10.1186/s12964-020-00603-z
Xu S, Ilyas I, Little PJ, Li H, Kamato D, Zheng X, Luo S, Li Z, Liu P, Han J, Harding IC, Ebong EE, Cameron SJ, Stewart AG, Weng J (2021) Endothelial dysfunction in atherosclerotic cardiovascular diseases and beyond: from mechanism to pharmacotherapies. Pharmacol Rev 73:924–967. https://doi.org/10.1124/pharmrev.120.000096
Zakkar M, Chaudhury H, Sandvik G, Enesa K, Le Luong A, Cuhlmann S, Mason JC, Krams R, Clark AR, Haskard DO, Evans PC (2008) Increased endothelial mitogen-activated protein kinase phosphatase-1 expression suppresses proinflammatory activation at sites that are resistant to atherosclerosis. Circ Res 103:726–732. https://doi.org/10.1161/circresaha.108.183913
Zhang Z, Zhang Q, Li F, Xin Y, Duan Z (2021) Contributions of HO-1-dependent MAPK to regulating intestinal barrier disruption. Biomol Ther (seoul) 29:175–183. https://doi.org/10.4062/biomolther.2020.112
Zheng X, Zhang W, Hu X (2018) Different concentrations of lipopolysaccharide regulate barrier function through the PI3K/Akt signalling pathway in human pulmonary microvascular endothelial cells. Sci Rep 8:9963. https://doi.org/10.1038/s41598-018-28089-3
Acknowledgements
This research was funded by National Research Foundation of Korea (KNRF-2019R1C1C1007331 and 2022R1A2C4001776).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
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.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Van Nguyen, D., Nguyen, T.L., Jin, Y. et al. 6′-Sialylactose abolished lipopolysaccharide-induced inflammation and hyper-permeability in endothelial cells. Arch. Pharm. Res. 45, 836–848 (2022). https://doi.org/10.1007/s12272-022-01415-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12272-022-01415-0