Pinacidil and levamisole prevent glutamate-induced death of hippocampal neuronal cells through reducing ROS production

Abstract

Activators of both adenosine 5'-triphosphate (ATP)-sensitive K(+) (KATP) channel and cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channel have significant in vivo and in vitro neuroprotection against glutamate-induced death of some neuronal cells. Here, the effect of the KATP channel activator, pinacidil, and the CFTR Cl(-) channel opener, levamisole, against glutamate-induced oxidative stress were investigated in mouse hippocampal cells, HT22. The results from cell viability assay (WST-1) showed that pinacidil and levamisole weakly protected cells against glutamate-induced toxicity at 10 μM and their effect increased in a dose-dependent manner till reach maximum protection at 300 μM. Pretreatment with pinacidil or levamisole significantly suppressed the elevation of reactive oxygen species (ROS) triggered by glutamate through stabilising mitochondrial membrane potential and subsequently protected HT22 cells against glutamate-induced death. HT22 cells viability was maintained by pinacidil and levamisole in presence of glutathione inhibitor, BSO. Also, pinacidil and levamisole pretreatment did not induce recovery of glutathione levels decreased by glutamate Expectedly, this protection was abolished by the KATP and CFTR Cl(-) channels blocker, glibenclamide. Thus, both pinacidil and levamisole protect HT22 cells against glutamate-induced cell death through stabilising mitochondrial membrane potential and subsequently decreasing ROS production.

Molecular Bases of Brain Preconditioning

“…As early as 3 h post-IP, we observed a decrease in levels of 3-nitroTyr (Figure 3D)—the marker of NO-dependent oxidative stress (Mohiuddin et al, 2006). The above indirectly points to the activation of the antioxidant systems as a result of tissue conditioning and is confirmed by the studies of neuronal survival, in which the pretreatment with diazoxide prevented cell death via antioxidative pathway activation (Virgili et al, 2013; Shukry et al, 2015). Indeed, diazoxide-induced mitochondrial membrane depolarization (Xi et al, 2005; Vadziuk et al, 2010) can lead to uncoupling of mitochondrial respiration and phosphorylation of adenosine diphosphate molecules (Holmuhamedov et al, 2004) and to the moderate production of ROS (Andrukhiv et al, 2006; Katakam et al, 2016), which may stimulate the antioxidant defense.…”

Huperzine A Alleviates Oxidative Glutamate Toxicity in Hippocampal HT22 Cells via Activating BDNF/TrkB-Dependent PI3K/Akt/mTOR Signaling Pathway

“…Actually, oxidative injury was previously reported to be triggered in HT22 cells under glutamate stimulation, and the medicinal herb Gastrodia elata remarkably prevented oxidative glutamate toxicity (Han et al 2014). Besides, several previous investigations illustrated that suppression of ROS production by various substances such as kaempferol (Yang et al 2014), acetogenin A (Lee et al 2015), pinacidil (Shukry et al 2015), or herbal mixture (Ahn et al 2015) could remarkably attenuate cell apoptosis in mouse hippocampal HT22 cells. Similarly, results from our current study illustrated that HupA pretreatment dramatically suppressed ROS accumulation and subsequent neuronal damage in HT22 cells.…”

Attenuated glutamate induced ROS production by antioxidative compounds in neural cell lines

“…The reactive oxygen species (ROS) triggered by glutamate was identify as a major reason for cells death. 36 Hence, we evaluated the relative ROS production by supplying with IC 50 concentration glutamate for each cell. The accumulation of ROS was measured by a typical way, utilizing a converting reaction of DCFH-DA to DCF.…”