PMID- 29141644 OWN - NLM STAT- MEDLINE DCOM- 20180625 LR - 20181113 IS - 1423-0127 (Electronic) IS - 1021-7770 (Print) IS - 1021-7770 (Linking) VI - 24 IP - 1 DP - 2017 Nov 15 TI - Lipopolysaccharide pretreatment increases protease-activated receptor-2 expression and monocyte chemoattractant protein-1 secretion in vascular endothelial cells. PG - 85 LID - 10.1186/s12929-017-0393-1 [doi] LID - 85 AB - BACKGROUND: This study investigated whether lipopolysaccharide (LPS) increase protease-activated receptor-2 (PAR-2) expression and enhance the association between PAR-2 expression and chemokine production in human vascular endothelial cells (ECs). METHODS: The morphology of ECs was observed through microphotography in cultured human umbilical vein ECs (EA. hy926 cells) treated with various LPS concentrations (0, 0.25, 0.5, 1, and 2 mug/mL) for 24 h, and cell viability was assessed using the MTT assay. Intracellular calcium imaging was performed to assess agonist (trypsin)-induced PAR-2 activity. Western blotting was used to explore the LPS-mediated signal transduction pathway and the expression of PAR-2 and adhesion molecule monocyte chemoattractant protein-1 (MCP-1) in ECs. RESULTS: Trypsin stimulation increased intracellular calcium release in ECs. The calcium influx was augmented in cells pretreated with a high LPS concentration (1 mug/mL). After 24 h treatment of LPS, no changes in ECs viability or morphology were observed. Western blotting revealed that LPS increased PAR-2 expression and enhanced trypsin-induced extracellular signal-regulated kinase (ERK)/p38 phosphorylation and MCP-1 secretion. However, pretreatment with selective ERK (PD98059), p38 mitogen-activated protein kinase (MAPK) (SB203580) inhibitors, and the selective PAR-2 antagonist (FSLLRY-NH2) blocked the effects of LPS-activated PAR-2 on MCP-1 secretion. CONCLUSIONS: Our findings provide the first evidence that the bacterial endotoxin LPS potentiates calcium mobilization and ERK/p38 MAPK pathway activation and leads to the secretion of the pro-inflammatory chemokine MCP-1 by inducing PAR-2 expression and its associated activity in vascular ECs. Therefore, PAR-2 exerts vascular inflammatory effects and plays an important role in bacterial infection-induced pathological responses. FAU - Chao, Hung-Hsing AU - Chao HH AD - Division of Cardiovascular Surgery, Department of Surgery, Shin Kong Wu Ho-Su Memorial Hospital, Taipei, 111, Taiwan. AD - Department of Surgery, School of Medicine, Taipei Medical University, Taipei, 11031, Taiwan. FAU - Chen, Po-Yuan AU - Chen PY AD - Department of Biological Science and Technology, College of Biopharmaceutical and Food Sciences, China Medical University, Taichung, 40402, Taiwan. FAU - Hao, Wen-Rui AU - Hao WR AD - Division of Cardiology, Department of Internal Medicine, Shuang Ho Hospital, Taipei Medical University, No. 291, Zhongzheng Rd, Zhonghe District, New Taipei City, 23561, Taiwan. FAU - Chiang, Wei-Ping AU - Chiang WP AD - Division of Cardiology, Department of Internal Medicine, Shuang Ho Hospital, Taipei Medical University, No. 291, Zhongzheng Rd, Zhonghe District, New Taipei City, 23561, Taiwan. FAU - Cheng, Tzu-Hurng AU - Cheng TH AD - Department of Biochemistry, School of Medicine, China Medical University, Taichung, 40402, Taiwan. AD - Department of Pharmacology & Graduate Institute of Pharmacology, National Defense Medical Center, Taipei, 114, Taiwan. FAU - Loh, Shih-Hurng AU - Loh SH AD - Department of Pharmacology & Graduate Institute of Pharmacology, National Defense Medical Center, Taipei, 114, Taiwan. FAU - Leung, Yuk-Man AU - Leung YM AD - Department of Physiology, School of Medicine, China Medical University, Taichung, 40402, Taiwan. FAU - Liu, Ju-Chi AU - Liu JC AD - Division of Cardiology, Department of Internal Medicine, Shuang Ho Hospital, Taipei Medical University, No. 291, Zhongzheng Rd, Zhonghe District, New Taipei City, 23561, Taiwan. AD - Department of Internal Medicine, School of Medicine, College of Medicine, Taipei Medical University, Taipei, 11031, Taiwan. FAU - Chen, Jin-Jer AU - Chen JJ AD - Graduate Institute of Clinical Medicine, College of Medicine, China Medical University, Taichung, 40402, Taiwan. AD - Institute of Biomedical Sciences, Academia Sinica, Taipei, 115, Taiwan. FAU - Sung, Li-Chin AU - Sung LC AUID- ORCID: 0000-0003-1564-5592 AD - Division of Cardiology, Department of Internal Medicine, Shuang Ho Hospital, Taipei Medical University, No. 291, Zhongzheng Rd, Zhonghe District, New Taipei City, 23561, Taiwan. 10204@s.tmu.edu.tw. AD - Department of Internal Medicine, School of Medicine, College of Medicine, Taipei Medical University, Taipei, 11031, Taiwan. 10204@s.tmu.edu.tw. LA - eng GR - SKH-8302-99-DR-15/Shin Kong Wu Ho-Su Memorial Hospital/ GR - SKH-TMU-101-07/Shin Kong Wu Ho-Su Memorial Hospital/ GR - TMU105-AE1-B21/Taipei Medical University/ PT - Journal Article DEP - 20171115 PL - England TA - J Biomed Sci JT - Journal of biomedical science JID - 9421567 RN - 0 (CCL2 protein, human) RN - 0 (Chemokine CCL2) RN - 0 (Lipopolysaccharides) RN - 0 (Receptor, PAR-2) SB - IM MH - Chemokine CCL2/*genetics/metabolism MH - Dose-Response Relationship, Drug MH - *Gene Expression MH - Human Umbilical Vein Endothelial Cells/*metabolism MH - Humans MH - Lipopolysaccharides/*pharmacology MH - Receptor, PAR-2/*genetics/metabolism MH - *Signal Transduction PMC - PMC5688698 OTO - NOTNLM OT - Endothelial cells OT - Lipopolysaccharides OT - Mitogen-activated protein kinases OT - Monocyte chemoattractant protein-1 OT - Protease-activated receptor-2 COIS- ETHICS APPROVAL AND CONSENT TO PARTICIPATE: Not applicable. CONSENT FOR PUBLICATION: All the authors have read and approved the paper for publication. COMPETING INTERESTS: The authors declare that they have no competing interests. PUBLISHER'S NOTE: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. EDAT- 2017/11/17 06:00 MHDA- 2018/06/26 06:00 PMCR- 2017/11/15 CRDT- 2017/11/17 06:00 PHST- 2017/05/21 00:00 [received] PHST- 2017/11/07 00:00 [accepted] PHST- 2017/11/17 06:00 [entrez] PHST- 2017/11/17 06:00 [pubmed] PHST- 2018/06/26 06:00 [medline] PHST- 2017/11/15 00:00 [pmc-release] AID - 10.1186/s12929-017-0393-1 [pii] AID - 393 [pii] AID - 10.1186/s12929-017-0393-1 [doi] PST - epublish SO - J Biomed Sci. 2017 Nov 15;24(1):85. doi: 10.1186/s12929-017-0393-1.