L-type voltage-gated calcium channels (L-VGCCs) are thought to be involved in epileptogenesis and acute excitotoxicity. However, little is known about the role of L-VGCCs in neuroinflammation or delayed neuronal death following excitotoxic insult. We examined the effects of repeated treatment with the L-VGCC blocker nimodipine on neuroinflammatory changes and delayed neuronal apoptosis in the dentate gyrus following trimethyltin (TMT)-induced convulsions. Male C57BL/6 N mice were administered TMT (2.6 mg/kg, i.p.), and the expression of the Cav1.2 and Cav1.3 subunits of L-VGCC were evaluated. The expression of both subunits was significantly decreased; however, the astroglial expression of Cav1.3 L-VGCC was significantly induced at 6 and 10 days after TMT treatment. Furthermore, astroglial Cav1.3 L-VGCCs colocalized with both the pro-inflammatory phenotype marker C3 and the anti-inflammatory phenotype marker S100A10 of astrocytes. Nimodipine (5 mg/kg, i.p. × 5 at 12-h intervals) did not significantly affect TMT-induced astroglial activation. However, nimodipine significantly attenuated the pro-inflammatory phenotype changes, while enhancing the anti-inflammatory phenotype changes in astrocytes after TMT treatment. Consistently, nimodipine reduced the levels of pro-inflammatory astrocytes-to-microglia mediators, while increasing the levels of anti-inflammatory astrocytes-to-microglia mediators. These effects were accompanied by an increase in the phosphorylation of extracellular signal-regulated kinase (ERK), supporting our previous finding that p-ERK is a signaling factor that regulates astroglial phenotype changes. In addition, nimodipine significantly attenuated TMT-induced microglial activation and delayed apoptosis of dentate granule neurons. Our results suggest that L-VGCC blockade attenuates neuroinflammation and delayed neurotoxicity following TMT-induced convulsions through the regulation of astroglial phenotypic changes by promoting ERK signaling.

OBJECTIVE: We here investigated the effect of late- and post-ictal treatment with rottlerin, a polyphenol compound isolated from Mallotus philippinensis, on delayed apoptotic neuronal death induced by trimethyltin (TMT) in mice.
METHODS: Male C57BL/6N mice received a single injection of TMT (2.4 mg/kg, i.p.), and mice were treated with rottlerin after a peak time (i.e., 2 d post-TMT) of convulsive behaviors and apoptotic cell death (5.0 mg/kg, i.p. at 3 and 4 d after TMT injection). Object location test and tail suspension test were performed at 5 d after TMT injection. In addition, changes in the expression of apoptotic and neurogenic markers in the dentate gyrus were examined.
RESULTS: Late- and post-ictal treatment with rottlerin suppressed delayed neuronal apoptosis in the dentate gyrus, and attenuated memory impairments (as evaluated by object location test) and depression-like behaviors (as evaluated by tail suspension test) at 5 days after TMT injection in mice. In addition, rottlerin enhanced the expression of Sox2 and DCX, and facilitated p-ERK expression in BrdU-incorporated cells in the dentate gyrus of TMT-treated mice. Rottlerin also increased p-Akt expression, and attenuated the increase in the ratio of pro-apoptotic factors/anti-apoptotic factors, and consequent cytosolic cytochrome c release and caspase-3 cleavage. Rottlerin-mediated action was significantly reversed by SL327, an ERK inhibitor.
CONCLUSIONS: Our results suggest that late- and post-ictal treatment with rottlerin attenuates TMT-induced delayed neuronal apoptosis in the dentate gyrus of mice via promotion of neurogenesis and inhibition of an on-going apoptotic process through up-regulation of p-ERK.