The role of extracellular Ca2+ in pancreatic acinar membrane damage (cellular injury) by nicotine, membrane-active agents (mellitin, snake venom and Ca2+ ionophore A23187) and secretagogues (CCK-8 and secretin) was investigated. Freshly isolated dispersed pancreatic acini from 18 h fasted adult rats were incubated with one of the aforementioned agents, in the absence and presence of Ca2+. Cellular injury was assessed by measuring the release of pulse-labeled 51Cr and LDH. In addition, release of amylase, trypsinogen and chymotrypsinogen was also determined. In the absence of Ca2+ nicotine (6 mM) caused a profound release of 51Cr and LDH as well as amylase, trypsinogen and chymotrypsinogen from the isolated pancreatic acini. Release of these enzymes and 51Cr decreased sharply with addition of increasing concentrations (0.25-5 mM) of Ca2+. Release of 51Cr and amylase by snake venom (50 micrograms/ml) was found to be 100 and 25% higher, respectively, in the absence of Ca2+ than in its presence. On the other hand, the Ca2+ ionophore A23187 (7 micrograms/ml) was found to be effective in releasing 51Cr and amylase only in the presence of Ca2+. CCK-8, (0.25nM), secretin (1 microM) and mellitin (0.5 microgram/ml) although significantly stimulated amylase secretion (225-350%) in the presence of Ca2+, none of the agents induced 51Cr release from acini, either in the absence or in the presence of extracellular Ca2+. It is concluded that the extracellular Ca2+ plays no specific role in cytotoxic injury in isolated pancreatic acini.
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