Extracellular and intracellular productions of lysophosphatidic acids and cyclic phosphatidic acids by lysophospholipase D from exogenously added lysophosphatidylcholines to cultured NRK52E cells |
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| Institution: | 1. Pharmaceutics, Graduate School of Clinical Pharmacy, Kyushu University of Health and Welfare, 1714-1 Yoshino-machi, Nobeoka, Miyazaki 882-8508, Japan;2. Faculty of Pharmacy, Yasuda Women''s University, Hiroshima, Japan;3. Graduate School of Biomedical Sciences, Tokushima University, Tokushima, Japan;4. Faculty of Pharmaceutical Sciences, Sojo University, Kumamoto, Japan;1. Servicio de Bioquímica Clínica, UCA-CCM, HU Ramón y Cajal-IRYCIS, Madrid, Spain;2. Instituto Murciano de Investigación Biosanitaria (IMIB-Arrixaca), Murcia, Spain;3. Servicio de Bioquímica-Investigación, HU Ramón y Cajal-IRYCIS, Madrid, Spain;4. Servicio de Gastroenterología, HU Ramón y Cajal-IRYCIS, Madrid, Spain;5. Servicio de Cirugía General, HCU Virgen de la Victoria, Málaga, Spain;6. Servicio de Anatomía Patológica, HU Ramón y Cajal-IRYCIS, Madrid, Spain;7. Departamento de Cirugía General y Aparato Digestivo, HU Virgen de la Arraixaca, Murcia, Spain;8. CIBER de Enfermedades Hepáticas y Digestivas (CIBEREHD), ISCIII, Spain;1. Department of Physiology, School of Basic Medical Sciences, Shenzhen University Medical School, Shenzhen University, Shenzhen 518055, Guangdong, China;2. GuangZhou Laboratory, No.9 XingDaoHuanBei Road, Guangzhou International Bio Island, Guangzhou 510005, Guangdong, China;1. Universidad Rey Juan Carlos, Dpto. de Ciencias Básicas de la Salud, Avda. de Atenas s/n. 28922, Alcorcón, Madrid, Spain;2. Institute of Biomedical Research “Alberto Sols” (CSIC-UAM), 28029 Madrid, Spain;3. Centro de Investigación Biomédica en Red de Diabetes y Enfermedades Metabólicas Asociadas (CIBER-dem), ISCIII, 28029 Madrid, Spain;4. MEMORISM Research Unit of University Rey Juan Carlos-Institute of Biomedical Research “Alberto Sols” (CSIC), Madrid, Spain;5. Bristol Renal, Translational Health Sciences, University of Bristol, Bristol, UK;1. Institute for Clinical Chemistry, University Hospital and University of Zürich, Zürich, Switzerland;2. Department of Neurology, The Third Xiangya Hospital, Central South University, Changsha, China;3. Institute for Biochemistry, University of Zürich; Zürich, Switzerland;4. National Clinical Research Center for Geriatric Disorders, Central South University, Changsha, China;1. Department of Medical Biotechnology and Translational Medicine, University of Milano, 20054 Segrate, Milano, Italy;2. UK Dementia Research Institute at UCL, London, UK;3. Neuro-Sys, 410 Chemin Départemental 60, 13120 Gardanne, France;4. Department of Immunology, St. Jude Children''s Research Hospital, Memphis, TN 38105, USA;5. Department of Chemistry, University of Milano, Milan, Italy |
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| Abstract: | Lysophosphatidic acid (LPA) is a bioactive lysophospholipid that is a notable biomarker of kidney injury. However, it is not clear how LPA is produced in renal cells. In this study, we explored LPA generation and its enzymatic pathway in a rat kidney-derived cell, NRK52E cells. Culturing of NRK52E cells with acyl lysophosphatidylcholine (acyl LPC), or lyso-platelet activating factor (lysoPAF, alkyl LPC) was resulted in increased extracellular level of choline, co-product with LPA by lysophospholipase D (lysoPLD). Their activities were enhanced by addition of calcium ions to the cell culture medium, but failed to be inhibited by S32826, an autotaxin (ATX)-specific inhibitor. Liquid chromatography-tandem mass spectrometric analysis revealed the small, but significant extracellular production of acyl LPA/cyclic phosphatidic acid (cPA) and alkyl LPA/cPA. The mRNA expression of glycerophosphodiesterase (GDE) 7 with lysoPLD activity was elevated in confluent NRK52E cells cultured over 3 days. GDE7 plasmid-transfection of NRK52E cells augmented both extracellular and intracellular productions of LPAs (acyl and alkyl) as well as extracellular productions of cPAs (acyl and alkyl) from exogenous LPCs (acyl and alkyl). These results suggest that intact NRK52E cells are able to produce choline and LPA/cPA from exogenous LPCs through the enzymatic action of GDE7 that is located on the plasma membranes and intracellular membranes. |
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