Cytoprotective effects of calbindin-d28k against antimycin-A induced hypoxic injury in proximal tubular cells

Ming Ju Wu, Li Wen Lai, Yeong Hau H. Lien

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

Intracellular calcium plays an important role on the pathogenesis of hypoxia-induced cellular injury. Calbindin-D28k, a cytosolic vitamin D-dependent calcium binding protein, can serve as a buffer to limit a surge in intracellular Ca2+ concentration ([Ca2+]i) induced by various stimulations. To evaluate the possible cytoprotective effect of calbindin-D28k against hypoxic injury in proximal tubular cells, a plasmid containing calbindin-D28k cDNA under the control of CMV immediate-early gene promoter was transfected into the murine proximal tubular epithelial (MCT) cells. The expression of calbindin-D28k in the transfected cells was verified with Northern blot analysis, Western blot analysis, and immunofluorescent staining. The non-transfected and transfected MCT cells were subjected to chemical hypoxia induced by antimycin A (10 μM) and glucose deprivation for 30-120 min. The transfection of calbindin-D28k reduced lactate dehydrogenase (LDH) release by 41%, 41%, 24%, and 24%, respectively, at 30, 60, 90 and 120 min after hypoxia when compared to the non-transfected cells (all p < 0.05). Cell viability after hypoxic injury was also significantly higher in transfected cells than non-transfected cells. Transfection with the plasmid without calbindin-D28k cDNA did not affect LDH release or cell viability after chemical hypoxic injury. [Ca+2]i was measured ratiometrically with fura-2 after exposure to chemical hypoxia. The rate of initial rise in [Ca2+]i and final [Ca+2]i at 30-120 min were significantly lowered in transfected cells. In conclusion, this study demonstrated that transfection of calbindin-D28k gene into MCT cells provide protective effects against chemical hypoxic injury probably through its buffering effects on [Ca+2]i.

Original languageEnglish (US)
Pages (from-to)559-569
Number of pages11
JournalLife Sciences
Volume71
Issue number5
DOIs
StatePublished - Jun 21 2002

Keywords

  • Intracellular calcium
  • Lactate dehydrogenase
  • Renal cells
  • Transfection

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Pharmacology, Toxicology and Pharmaceutics(all)

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