NPC-14686, a novel anti-inflammatory agent, increased intracellular Ca2+ concentrations in MDCK renal tubular cells

Chung Ren Jan*, Jue Long Wang, Kang Ju Chou, Jin Shiung Cheng, Kam Chung Lee, Li Ling Tseng, Shiou Ping Wang, Kwong Yui Tang, Jong Khing Huang

*Corresponding author for this work

Research output: Contribution to journalJournal Article peer-review

10 Scopus citations

Abstract

The effect of NPC-14686 (Fmoc-L-homophenylalanine), a novel anti-inflammatory agent on intracellular free Ca2+ concentrations ([Ca2+](i)) in Madin Darby canine kidney (MDCK) renal tubular cells, was investigated, using fura-2 as a Ca2+ dye. At concentrations between 10 and 200 μM NPC-14686 increased [Ca2+](i) concentration dependently. The [Ca2+](i) signal comprised an initial rise and a sustained phase. Ca2+ removal inhibited the Ca2+ signals by 90%. In Ca2+-free medium, pretreatment with 100 μM NPC-14686 nearly abolished the [Ca2+](i) increase induced by 1 μM thapsigargin (an endoplasmic reticulum Ca2+ pump inhibitor) and abolished the [Ca2+](i) increase induced by 2 μM carbonylcyanide m-chlorophenylhydrazone (CCCP) (a mitochondrial uncoupler). NPC-14686 (100 μM) induced a slight [Ca2+](i) increase after pretreatment with 2 μM CCCP and 1 μM thapsigargin. Addition of 3 mM Ca2+ elicited a [Ca2+](i) increase in cells pretreated with 100 μM NPC-14686 in Ca2+-free medium. Inhibition of inositol-1,4,5-trisphosphate (IP3) production by suppressing phospholipase C with 2 μM U73122 did not alter NPC-14686-induced Ca2+ release. Trypan blue exclusion revealed that incubation with 10 or 200 μM NPC-14686 for 1-30 min decreased cell viability by 10-20% concentration dependently. Collectively, the results demonstrate that, in MDCK tubular cells, NPC-14686 induced Ca2+ release followed by Ca2+ entry, with the latter playing a major role. NPC-14686 appears to release intracellular Ca2+ in an IP3-uncoupled manner. NPC-14686 may be of mild cytotoxicity. Copyright (C) 2000 International Society for Immunopharmacology. Published by Elsevier Science Ltd.

Original languageEnglish
Pages (from-to)915-921
Number of pages7
JournalInternational Journal of Immunopharmacology
Volume22
Issue number11
DOIs
StatePublished - 2000
Externally publishedYes

Keywords

  • Fura-2
  • Intracellular signaling
  • MDCK cells
  • NPC-14686
  • Renal pharmacology

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