Toxicity and physiological actions of biflavones on potassium current in insect neuronal cells.

ABSTRACT

Stellera chamaejasme L. is a Chinese traditional herb. It has a long history and many medicinal usages. Biflavones, one of the main active ingredients in S. chamaejasme's roots, possess excellent insecticidal activities both in vivo and in vitro. However, the mechanism of these compounds and its potential molecular targets on insect cell were still not clear. Here the whole cell patch clamp technique was used to investigate whether biflavones affects voltage-gated potassium channels (Kv) on insect neuronal cells (AW1 and WG2). The results confirmed that both the three biflavones neochamaejasmin A (NCA), neochamaejasmin B (NCB) and isochamaejasmin A (ICM) can significantly inhibit the A-type potassium current (IA) than delayed rectifier potassium current (IK) expressed on insect cells. Moreover, ICM stood out as the strongest inhibition activity on IA with IC50 value of 106.75 µM. Multiple results suggest that the inhibition of potassium current was related to the gating modification of biflavones. ICM produced concentration dependent hyperpolarizing shifts in the voltage dependence of channel steady-state activation and inactivation. Maximal shifts of the ICM-induced V0.5, were -15.1 mV for activation and -6.93 mV for inactivation. ICM also prolonged recovery from inactivation of current. Moreover, the biflavones could inhibited AW1 cell survival in both dose- and time-dependent manners with well correlation of K+ inhibitory activity. Our study showed that biflavones from S. chamaejasme exhibiting significant blocked effects on Kv of AW1 cells and inhibited cell proliferation. These findings may not only show the toxic mechanisms of biflavones on insect cells, but also suggest that Kv channel play an important role in biflavones' mode of action and may be the new targets for designing novel insecticides.