Justicidin A Reduces β-Amyloid via Inhibiting Endocytosis of β-Amyloid Precursor Protein

β-amyloid precursor protein (APP) can be cleaved by α-, and γ-secretase at plasma membrane producing soluble ectodomain fragment (sAPPα). Alternatively, following endocytosis, APP is cleaved by β-, and γ-secretase at early endosomes generating β-amyloid (Aβ), the main culprit in Alzheimer's disease (AD). Thus, APP endocytosis is critical for Aβ production. Recently, we reported that Monsonia angustifolia, the indigenous vegetables consumed in Tanzania, improved cognitive function and decreased Aβ production. In this study, we examined the underlying mechanism of justicidin A, the active compound of M. angustifolia, on Aβ production. We found that justicidin A reduced endocytosis of APP, increasing sAPPα level, while decreasing Aβ level in HeLa cells overexpressing human APP with the Swedish mutation. The effect of justicidin A on Aβ production was blocked by endocytosis inhibitors, indicating that the decreased APP endocytosis by justicidin A is the underlying mechanism. Thus, justicidin A, the active compound of M. angustifolia, may be a novel agent for AD treatment.

Effect of capsaicin on delayed rectifier K+ current in adult rat dorsal root ganglion neurons

K+ currents play multiple roles in the excitability of dorsal root ganglion (DRG) neurons. Influences on these currents change the shape of the action potential, its firing threshold and the resting membrane potential. In this study, whole cell configuration of patch clamp technique had been applied to record the blocking effect of capsaicin, a lipophilic alkaloid, on the delayed rectifier K+ current in cultured small diameter DRG neurons of adult rat. Capsaicin reduced the amplitude of K+ current in dose dependent manner, and the concentration-dependence curve was well described by the Hill equation with KD value of 19.1 micrometer. The blocking effect of capsaicin was reversible. Capsaicin (10 micrometer) shifted the steady-state inactivation curve in the hyperpolarizing direction by about 15 mV and increased the rate of inactivation. The voltage dependence of activation was not affected by capsaicin. These multiple effects of capsaicin may suggest that capsaicin bind to the region of K+ channel, participating in inactivation process.

External pH effects on delayed rectifier K+ currents of small dorsal root ganglion neuron of rat

Under certain pathophysiological conditions, such as inflammation and ischemia, the concentration of H+ ion in the tissue surrounding neurons is changed. Variations in H+ concentration are known to alter the conduction and/of the gating properties of several types of ion channels. Several types of K+ channels are modulated by pH. In this study, the whole cell configuration of the patch clamp technique has been applied to the recording of the responses of change of external pH on the delayed rectifier K+ current of cultured DRG neurons of rat. Outward K+ currents were examined in DRG cells, and the Charybdotoxin and Mn2+ could eliminate Ca2+-dependent K+ currents from outward K+ currents. This outward K+ current was activated around -60 mV by step depolarizing pulses from holding potential -70 mV. Outward K+ currents were decreased by low external pH. Activation and steady-state inactivation curve were shifted to the right by acidification, while there was small change by alkalization. These results suggest that H+ could be alter the sensory modality by changing and modifying voltage-dependent K+ currents, which participated in repolarization.

Dual action of d-tubocurarine on large-conductance Ca2+-activated K+ channels from rat brain reconstituted into planar lipid bilayer

Using the planar lipid bilayer method, we investigated the effect of d-tubocurarine (dTC) on the extracellular side of large-conductance Ca2+-activated K+ channel from rat brain. When the initial open probability (Po) of the channel was relatively high, dTC decreased channel activity in a concentration dependent manner. In contrast, when the initial Po was lower, sub-micro molar dTC increased channel activity by destabilizing the closed states of the channel. Further addition of dTC up to micro molar range decreased channel activity. This dual effect of dTC implicates that there exist at least two different binding sites for dTC.