OLeandrin Produces Changes in Intracellular Calcium Levels in Isolated Cardiomyocytes: A Real-Time Fluorescence Imaging Study Comparing Adult to Neonatal Cardiomyocytes

Abstract

Oleanders are common, hardy shrubs that grow throughout the southern United States. They contain cardiotonic steroids formed from cardenolides and bufadienolides, making the plant poisonous to both animals and humans. Aliquots of both commercially available oleander and fresh oleander extracts were prepared. Fresh, rod-like, calcium-tolerant adult rat cardiomyocytes and cultured neonatal cardiomyocytes were isolated and treated with 0–4 ng/ml of both preparations, challenged with verapamil and ouabain, and real-time spectrophotometric calcium transients and images were acquired. A number of effects were observed with the adult cells: (1) intracellular calcium levels were increased in a concentration-dependent manner: (2) reduced calcium transient heights and eventual cessation of beating resulted; and (3) increased sparking intensity led to subsequent beating and eventual calcium overload. In the spontaneously beating cultured neonatal myocytes increased intramyocytic calcium levels were also seen, with retention of this calcium rise leading to overload and, as in the adult myocytes, cessation of beating. These observations demonstrate that oleander extract is markedly potent with respect to the elevation of calcium concentrations in cardiomyocytes, and that the inability of the cardiomyocytes to release the accumulated calcium possibly indicates a role for oleandrin in inhibition of ryanodine receptor calcium release channels, calcium uptake via Na⁺,K⁺-ATPase inhibition [EC 3.6.1.3], and/or dysfunction of sarcolemmal calcium release channels.