Biomolecules & Therapeutics

The Korean Society of Applied Pharmacology (한국응용약물학회)
권호 표지
DOI QR코드

DOI QR Code

Thrombin Induced Apoptosis through Calcium-Mediated Activation of Cytosolic Phospholipase A2 in Intestinal Myofibroblasts

  • Mi Ja, Park (Department of Environmental & Health Chemistry, College of Pharmacy, Chung-Ang University) ;
  • Jong Hoon, Won (Department of Environmental & Health Chemistry, College of Pharmacy, Chung-Ang University) ;
  • Dae Kyong, Kim (Department of Environmental & Health Chemistry, College of Pharmacy, Chung-Ang University)
  • Received : 2022.05.25
  • Accepted : 2022.07.06
  • Published : 2023.01.01
Download PDF
⟨ Previous Next ⟩

Abstract

Thrombin is a serine protease that participates in a variety of biological signaling through protease-activated receptors. Intestinal myofibroblasts play central roles in maintaining intestinal homeostasis. In this study, we found that thrombin-induced apoptosis is mediated by the calcium-mediated activation of cytosolic phospholipase A2 in the CCD-18Co cell. Thrombin reduced cell viability by inducing apoptosis and proteinase-activated receptor-1 antagonist attenuated thrombin-induced cell death. Endogenous ceramide did not affect the cell viability itself, but a ceramide-mediated pathway was involved in thrombin-induced cell death. Thrombin increased intracellular calcium levels and cytosolic phospholipase A2 activity. The ceramide synthase inhibitor Fumonisin B1, intracellular calcium chelator BAPTA-AM, and cytosolic phospholipase A2 inhibitor AACOCF3 inhibited thrombin-induced cell death. Thrombin stimulated arachidonic acid release and reactive oxygen species generation, which was blocked by AACOCF3, BAPTA-AM, and the antioxidant reagent Trolox. Taken together, thrombin triggered apoptosis through calcium-mediated activation of cytosolic phospholipase A2 in intestinal myofibroblasts.

Keywords

Acknowledgement

This study was supported by grants from the National Research Foundation of Korea (NRF-2017M3A9D8048414) funded by the Korean government (Ministry of Science and ICT).

References

  1. Ahmad, R., Knafo, L., Xu, J., Sindhu, S. T., Menezes, J. and Ahmad, A. (2000) Thrombin induces apoptosis in human tumor cells. Int. J. Cancer 87, 707-715. https://doi.org/10.1002/1097-0215(20000901)87:5<707::AID-IJC13>3.0.CO;2-W
  2. Anrather, D., Millan, M. T., Palmetshofer, A., Robson, S. C., Geczy, C., Ritchie, A. J., Bach, F. H. and Ewenstein, B. M. (1997) Thrombin activates nuclear factor-kappaB and potentiates endothelial cell activation by TNF. J. Immunol. 159, 5620-5628. https://doi.org/10.4049/jimmunol.159.11.5620
  3. Beltinger, J., Mckaig, B. C., Makh, S., Stack, W. A., Hawkey, C. J. and Mahida, Y. R. (1999) Human colonic subepithelial myofibroblasts modulate transepithelial resistance and secretory response. Am. J. Physiol. 277, C271-C279. https://doi.org/10.1152/ajpcell.1999.277.2.C271
  4. Bhalla, D. K., Hirata, F., Rishi, A. K. and Gairola, C. G. (2009) Cigarette smoke, inflammation, and lung injury: a mechanistic perspective. J. Toxicol. Environ. Health B Crit. Rev. 12, 45-64. https://doi.org/10.1080/10937400802545094
  5. Chande, N., Mcdonald, J. W. and Macdonald, J. K. (2008) Unfractionated or low-molecular weight heparin for induction of remission in ulcerative colitis. Cochrane Database Syst. Rev. (2), CD006774.
  6. Chin, A. C., Vergnolle, N., Macnaughton, W. K., Wallace, J. L., Hollenberg, M. D. and Buret, A. G. (2003) Proteinase-activated receptor 1 activation induces epithelial apoptosis and increases intestinal permeability. Proc. Natl. Acad. Sci. U.S.A. 100, 11104-11109. https://doi.org/10.1073/pnas.1831452100
  7. Choi, J. M., Choi, Y. H., Kim, S. K., Ahn, K. H., Won, J. H., Lim, J. H., Jang, Y. J., Lee, S., Kim, D. H. and Kim, D. K. (2013) (S)-tetrahydroisoquinoline alkaloid inhibits LPS-induced arachidonic acid release through downregulation of cPLA2 expression. Mol. Cells 36, 400-409. https://doi.org/10.1007/s10059-013-0078-x
  8. Corvera, C. U., Dery, O., Mcconalogue, K., Gamp, P., Thoma, M., Al-Ani, B., Caughey, G. H., Hollenberg, M. D. and Bunnett, N. W. (1999) Thrombin and mast cell tryptase regulate guinea-pig myenteric neurons through proteinase-activated receptors-1 and -2. J. Physiol. 517, 741-756. https://doi.org/10.1111/j.1469-7793.1999.0741s.x
  9. Denadai-Souza, A., Bonnart, C., Tapias, N. S., Marcellin, M., Gilmore, B., Alric, L., Bonnet, D., Burlet-Schiltz, O., Hollenberg, M. D., Vergnolle, N. and Deraison, C. (2018) Functional proteomic profiling of secreted serine proteases in health and inflammatory bowel disease. Sci. Rep. 8, 7834.
  10. Donovan, F. M., Pike, C. J., Cotman, C. W. and Cunningham, D. D. (1997) Thrombin induces apoptosis in cultured neurons and astrocytes via a pathway requiring tyrosine kinase and RhoA activities. J. Neurosci. 17, 5316-5326. https://doi.org/10.1523/jneurosci.17-14-05316.1997
  11. Flynn, A. N. and Buret, A. G. (2004) Proteinase-activated receptor 1 (PAR-1) and cell apoptosis. Apoptosis 9, 729-737. https://doi.org/10.1023/B:APPT.0000045784.49886.96
  12. Horiguchi, H., Endo, M., Kawane, K., Kadomatsu, T., Terada, K., Morinaga, J., Araki, K., Miyata, K. and Oike, Y. (2017) ANGPTL2 expression in the intestinal stem cell niche controls epithelial regeneration and homeostasis. EMBO J. 36, 409-424. https://doi.org/10.15252/embj.201695690
  13. Jang, Y. J., Won, J. H., Back, M. J., Fu, Z., Jang, J. M., Ha, H. C., Hong, S., Chang, M. and Kim, D. K. (2015) Paraquat induces apoptosis through a mitochondria-dependent pathway in RAW264.7 cells. Biomol. Ther. (Seoul) 23, 407-413. https://doi.org/10.4062/biomolther.2015.075
  14. Karakhanova, S., Golovastova, M., Philippov, P. P., Werner, J. and Bazhin, A. V. (2014) Interlude of cGMP and cGMP/protein kinase G type 1 in pancreatic adenocarcinoma cells. Pancreas 43, 784-794. https://doi.org/10.1097/MPA.0000000000000104
  15. Kohjimoto, Y., Kennington, L., Scheid, C. R. and Honeyman, T. W. (1999) Role of phospholipase A2 in the cytotoxic effects of oxalate in cultured renal epithelial cells. Kidney Int. 56, 1432-1441. https://doi.org/10.1046/j.1523-1755.1999.00683.x
  16. Kolesnick, R. and Hannun, Y. A. (1999) Ceramide and apoptosis. Trends Biochem. Sci. 24, 224-225; author reply 227. https://doi.org/10.1016/S0968-0004(99)01408-5
  17. Kolesnick, R. N. and Hemer, M. R. (1990) Characterization of a ceramide kinase activity from human leukemia (HL-60) cells. Separation from diacylglycerol kinase activity. J. Biol. Chem. 265, 18803-18808. https://doi.org/10.1016/S0021-9258(17)30584-7
  18. Korbecki, J., Baranowska-Bosiacka, I., Gutowska, I. and Chlubek, D. (2013) The effect of reactive oxygen species on the synthesis of prostanoids from arachidonic acid. J. Physiol. Pharmacol. 64, 409-421.
  19. Kriem, B., Sponne, I., Fifre, A., Malaplate-Armand, C., Lozac'h-Pillot, K., Koziel, V., Yen-Potin, F. T., Bihain, B., Oster, T., Olivier, J. L. and Pillot, T. (2005) Cytosolic phospholipase A2 mediates neuronal apoptosis induced by soluble oligomers of the amyloid-beta peptide. FASEB J. 19, 85-87. https://doi.org/10.1096/fj.04-1807fje
  20. Li, G., Petiwala, S. M., Yan, M., Won, J. H., Petukhov, P. A. and Johnson, J. J. (2016) Gartanin, an isoprenylated xanthone from the mangosteen fruit (Garcinia mangostana), is an androgen receptor degradation enhancer. Mol. Nutr. Food Res. 60, 1458-1469. https://doi.org/10.1002/mnfr.201600037
  21. Messier, E. M., Bahmed, K., Tuder, R. M., Chu, H. W., Bowler, R. P. and Kosmider, B. (2013) Trolox contributes to Nrf2-mediated protection of human and murine primary alveolar type II cells from injury by cigarette smoke. Cell Death Dis. 4, e573.
  22. Meyer, M. and Jaspers, I. (2015) Respiratory protease/antiprotease balance determines susceptibility to viral infection and can be modified by nutritional antioxidants. Am. J. Physiol. Lung Cell. Mol. Physiol. 308, L1189-L1201. https://doi.org/10.1152/ajplung.00028.2015
  23. Motta, J. P., Denadai-Souza, A., Sagnat, D., Guiraud, L., Edir, A., Bonnart, C., Sebbag, M., Rousset, P., Lapeyre, A., Seguy, C., Mathurine-Thomas, N., Galipeau, H. J., Bonnet, D., Alric, L., Buret, A. G., Wallace, J. L., Dufour, A., Verdu, E. F., Hollenberg, M. D., Oswald, E., Serino, M., Deraison, C. and Vergnolle, N. (2019) Active thrombin produced by the intestinal epithelium controls mucosal biofilms. Nat. Commun. 10, 3224.
  24. Motta, J. P., Deraison, C., Le Grand, S., Le Grand, B. and Vergnolle, N. (2021a) PAR-1 antagonism to promote gut mucosa healing in Crohn's disease patients: a new avenue for CVT120165. Inflamm. Bowel Dis. 27, S33-S37. https://doi.org/10.1093/ibd/izab244
  25. Motta, J. P., Magne, L., Descamps, D., Rolland, C., Squarzoni-Dale, C., Rousset, P., Martin, L., Cenac, N., Balloy, V., Huerre, M., Frohlich, L. F., Jenne, D., Wartelle, J., Belaaouaj, A., Mas, E., Vinel, J. P., Alric, L., Chignard, M., Vergnolle, N. and Sallenave, J. M. (2011) Modifying the protease, antiprotease pattern by elafin overexpression protects mice from colitis. Gastroenterology 140, 1272-1282. https://doi.org/10.1053/j.gastro.2010.12.050
  26. Motta, J. P., Palese, S., Giorgio, C., Chapman, K., Denadai-Souza, A., Rousset, P., Sagnat, D., Guiraud, L., Edir, A., Seguy, C., Alric, L., Bonnet, D., Bournet, B., Buscail, L., Gilletta, C., Buret, A. G., Wallace, J. L., Hollenberg, M. D., Oswald, E., Barocelli, E., Le Grand, S., Le Grand, B., Deraison, C. and Vergnolle, N. (2021b) Increased mucosal thrombin is associated with Crohn's disease and causes inflammatory damage through protease-activated receptors activation. J. Crohns Colitis 15, 787-799. https://doi.org/10.1093/ecco-jcc/jjaa229
  27. Pettus, B. J., Bielawska, A., Subramanian, P., Wijesinghe, D. S., Maceyka, M., Leslie, C. C., Evans, J. H., Freiberg, J., Roddy, P., Hannun, Y. A. and Chalfant, C. E. (2004) Ceramide 1-phosphate is a direct activator of cytosolic phospholipase A2. J. Biol. Chem. 279, 11320-11326. https://doi.org/10.1074/jbc.M309262200
  28. Pettus, B. J., Chalfant, C. E. and Hannun, Y. A. (2002) Ceramide in apoptosis: an overview and current perspectives. Biochim. Biophys. Acta 1585, 114-125. https://doi.org/10.1016/S1388-1981(02)00331-1
  29. Pinton, P., Giorgi, C., Siviero, R., Zecchini, E. and Rizzuto, R. (2008) Calcium and apoptosis: ER-mitochondria Ca2+ transfer in the control of apoptosis. Oncogene 27, 6407-6418. https://doi.org/10.1038/onc.2008.308
  30. Pompeia, C., Freitas, J. J., Kim, J. S., Zyngier, S. B. and Curi, R. (2002) Arachidonic acid cytotoxicity in leukocytes: implications of oxidative stress and eicosanoid synthesis. Biol. Cell 94, 251-265. https://doi.org/10.1016/S0248-4900(02)01200-5
  31. Powell, D. W., Mifflin, R. C., Valentich, J. D., Crowe, S. E., Saada, J. I. and West, A. B. (1999) Myofibroblasts. II. Intestinal subepithelial myofibroblasts. Am. J. Physiol. 277, C183-C201. https://doi.org/10.1152/ajpcell.1999.277.2.C183
  32. Powell, D. W., Pinchuk, I. V., Saada, J. I., Chen, X. and Mifflin, R. C. (2011) Mesenchymal cells of the intestinal lamina propria. Annu. Rev. Physiol. 73, 213-237. https://doi.org/10.1146/annurev.physiol.70.113006.100646
  33. Rezaie, A. R. (2014) Protease-activated receptor signalling by coagulation proteases in endothelial cells. Thromb. Haemost. 112, 876-882. https://doi.org/10.1160/th14-02-0167
  34. Rukoyatkina, N., Mindukshev, I., Walter, U. and Gambaryan, S. (2013) Dual role of the p38 MAPK/cPLA2 pathway in the regulation of platelet apoptosis induced by ABT-737 and strong platelet agonists. Cell Death Dis. 4, e931.
  35. Sacks, R. S., Firth, A. L., Remillard, C. V., Agange, N., Yau, J., Ko, E. A. and Yuan, J. X. (2008) Thrombin-mediated increases in cytosolic [Ca2+] involve different mechanisms in human pulmonary artery smooth muscle and endothelial cells. Am. J. Physiol. Lung Cell. Mol. Physiol. 295, L1048-L1055. https://doi.org/10.1152/ajplung.90259.2008
  36. Scaldaferri, F., Lancellotti, S., Pizzoferrato, M. and De Cristofaro, R. (2011) Haemostatic system in inflammatory bowel diseases: new players in gut inflammation. World J. Gastroenterol. 17, 594-608. https://doi.org/10.3748/wjg.v17.i5.594
  37. Seymour, M. L., Zaidi, N. F., Hollenberg, M. D. and Macnaughton, W. K. (2003) PAR1-dependent and independent increases in COX-2 and PGE2 in human colonic myofibroblasts stimulated by thrombin. Am. J. Physiol. Cell Physiol. 284, C1185-C1192. https://doi.org/10.1152/ajpcell.00126.2002
  38. Simon, H. U., Haj-Yehia, A. and Levi-Schaffer, F. (2000) Role of reactive oxygen species (ROS) in apoptosis induction. Apoptosis 5, 415-418. https://doi.org/10.1023/a:1009616228304
  39. Smith, P. K., Krohn, R. I., Hermanson, G. T., Mallia, A. K., Gartner, F. H., Provenzano, M. D., Fujimoto, E. K., Goeke, N. M., Olson, B. J. and Klenk, D. C. (1985) Measurement of protein using bicinchoninic acid. Anal. Biochem. 150, 76-85. https://doi.org/10.1016/0003-2697(85)90442-7
  40. Smith, W. L. (1992) Prostanoid biosynthesis and mechanisms of action. Am. J. Physiol. 263, F181-F191. https://doi.org/10.1152/ajprenal.1992.263.2.F181
  41. Suzuki, A., Kozawa, O., Shinoda, J., Watanabe-Tomita, Y., Saito, H. and Oiso, Y. (1997) Mechanism of thrombin-induced arachidonic acid release in osteoblast-like cells. Prostaglandins Leukot. Essent. Fatty Acids 56, 467-472. https://doi.org/10.1016/S0952-3278(97)90601-3
  42. Turgeon, V. L., Lloyd, E. D., Wang, S., Festoff, B. W. and Houenou, L. J. (1998) Thrombin perturbs neurite outgrowth and induces apoptotic cell death in enriched chick spinal motoneuron cultures through caspase activation. J. Neurosci. 18, 6882-6891. https://doi.org/10.1523/jneurosci.18-17-06882.1998
  43. Valentich, J. D., Popov, V., Saada, J. I. and Powell, D. W. (1997) Phenotypic characterization of an intestinal subepithelial myofibroblast cell line. Am. J. Physiol. 272, C1513-C1524. https://doi.org/10.1152/ajpcell.1997.272.5.C1513
  44. Vergnolle, N. (2004) Modulation of visceral pain and inflammation by protease-activated receptors. Br. J. Pharmacol. 141, 1264-1274. https://doi.org/10.1038/sj.bjp.0705750
  45. Vergnolle, N. (2016) Protease inhibition as new therapeutic strategy for GI diseases. Gut 65, 1215-1224. https://doi.org/10.1136/gutjnl-2015-309147
  46. Walsh, T. G., Berndt, M. C., Carrim, N., Cowman, J., Kenny, D. and Metharom, P. (2014) The role of Nox1 and Nox2 in GPVI-dependent platelet activation and thrombus formation. Redox. Biol. 2, 178-186. https://doi.org/10.1016/j.redox.2013.12.023
  47. Won, J. H., Jeon, H. J., Kim, S. K., Shin, I. C., Jang, J. M., Ha, H. C., Back, M. J. and Kim, D. K. (2020) Interaction of synaptosomalassociated protein 25 with neutral sphingomyelinase 2: functional impact on the sphingomyelin pathway. Neuroscience 427, 1-15. https://doi.org/10.1016/j.neuroscience.2019.08.015
  48. Zain, J., Huang, Y. Q., Feng, X., Nierodzik, M. L., Li, J. J. and Karpatkin, S. (2000) Concentration-dependent dual effect of thrombin on impaired growth/apoptosis or mitogenesis in tumor cells. Blood 95, 3133-3138. https://doi.org/10.1182/blood.v95.10.3133.010k31_3133_3138
  49. Zhu, Y., Hua, P. and Lance, P. (2003) Cyclooxygenase-2 expression and prostanoid biogenesis reflect clinical phenotype in human colorectal fibroblast strains. Cancer Res. 63, 522-526.