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Anti-botulism triterpenoid toosendanin elicits calcium increase and exocytosis in rat sensory neurons

【摘要】:正Toosendanin,a triterpenoid from Melia toosendan Sieb et Zucc,has been found before to be an effective anti-botulism agent,with a bi-phasic effect at both motor nerve endings and central synapse;an initial facilitation followed by prolonged depression. Initial facilitation may be due to activation of voltage-dependent calcium channels plus inhibition of potassium channels,but the depres- sion is not fully understood.Toosendanin has no effect on intracellular calcium or secretion in the non-excitable pancreatic acinar cells, ruling out general toosendanin inhibition of exocytosis.In the present work toosendanin effects on sensory neurons isolated from rat nodose ganglia were investigated.It was found that toosendanin stimulated increases in cytosolic calcium and neuronal exocytosis dose-dependently. Experiments with membrane potential indicator bis-(1,3-dibutylbarbituric acid) trimethine oxonol found that toosendanin hyperpolarized capsaicin-insensitive but depolarized capsaicin-sensitive neurons;high potassium-induced calcium increase was much smaller in hyperpolarizing neurons than in depolarizing neurons,whereas no difference was found for potassium-induced depolarization in these two types of neurons.In neurons showing spontaneous calcium oscillations,toosendanin increased the oscillatory amplitude but not frequency.Toosendanin-induced calcium increase was decreased in calcium-free buffer,by nifedipine,and by transient receptor potential vanilloid 1(TRPV1) antagonist capsazepine.Simultaneous measurements of cytosolic and endoplasmic reticulum(ER) calcium showed an increase in cytosolic but a decrease in ER calcium,indicating that toosendanin triggered ER calcium release.These data together indicate that toosendanin modulates sensory neurons,but had opposite effects on membrane potential depending on the presence or absence of capsaicin receptor/TRPV 1 channel.

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