Release of acetylcholine to raise insulin secretion in Wistar rats by oleanolic acid, one of the active principles contained in Cornus officinalis.

The plasma glucose lowering action of fruits of cornus (Cornus officinalis), the major active constituent of Die-Huang-Wan, has been documented to mediate acetylcholine (ACh) release, which in turn to stimulate muscarinic M(3) receptors resulting in the enhancement of insulin secretion in rats with functional pancreatic beta-cells. The present study was conducted to investigate the effect of oleanolic acid, one of the active principles of cornus fruit, on the release of insulin in rats. After an intraperitoneal injection into the fasting Wistar rats for 90 min, oleanolic acid decreased the plasma glucose in a dose-dependent manner in parallel to an increase of plasma levels of insulin as well as C-peptide. Moreover, disruption of synaptic ACh using an inhibitor of choline uptake, hemicholinium-3, or vesicular acetylcholine transport, vesamicol, abolished these actions of oleanolic acid. Also, physostigmine at concentration sufficient to inhibit acetylcholinesterase enhanced the actions of oleanolic acid. Both the plasma glucose lowering action and the raised plasma levels of insulin and C-peptide induced by oleanolic acid were also inhibited by 4-diphenylacetoxy-N-methylpiperdine methiodide (4-DAMP), but not affected by the ganglionic nicotinic antagonist, pentolinium or hexamethonium. The results suggest that oleanolic acid has an ability to raise the release of ACh from nerve terminals, which in turn to stimulate muscarinic M(3) receptors in the pancreatic cells and augment the insulin release to result in plasma glucose lowering action. Thus, oleanolic acid is one of the active principles responsible for the increase of plasma insulin produced by cornus fruit in rats.