Background: Epilepsy is a chronic neurological disorder characterized by the recurrence of seizures. One-third of patients with epilepsy may not respond to antiseizure drugs. Purpose: We aimed to examine whether D-limonene, a cyclic monoterpene, exhibited any antiseizure activity in the pentylenetetrazole (PTZ)-induced kindling mouse model and in vitro. Methods: PTZ kindling mouse model was established by administering PTZ (30 mg/kg) intraperitoneally to mice once every 48 h. We performed immunoblot blots, immunohistochemistry (IHC), and high-performance liquid chromatography (HPLC) analysis after the behavioral study. Results: An acute injection of PTZ (60 mg/kg) induced seizure in mice, while pretreatment with D-limonene inhibited PTZ-induced seizure. Repeated administration of PTZ (30 mg/kg) increased the seizure score gradually in mice, which was reduced in D-limonene (10 mg/kg)-pretreated group. In addition, D-limonene treatment increased glutamate decarboxylase-67 (GAD-67) expression in the hippocampus. Axonal sprouting of hippocampal neurons after kindling was inhibited by D-limonene pretreatment. Moreover, D-limonene reduced the expression levels of Neuronal PAS Domain Protein 4 (Npas4)-induced by PTZ. Furthermore, the adenosine A2A antagonist SCH58261 and ZM241385 inhibited anticonvulsant activity and gamma-aminobutyric acid (GABA)ergic neurotransmission-induced by D-limonene. Conclusion: These results suggest that D-limonene exhibits anticonvulsant activity through modulation of adenosine A2A receptors on GABAergic neuronal function.
Even though synthetic colorants can cause side effects such as allergies and pigmentation, they have not been sufficiently researched. Herein, high-performance liquid chromatography, liquid chromatography-tandem mass spectrometry, and liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS) were used to detect 13 banned synthetic colorants in cosmetics and characterize their fragmentation. The developed HPLC method was validated following the International Conference on Harmonisation guidelines (specificity, limit of detection, limit of quantification, recovery, linearity, accuracy, and precision) and applied to 120 distributed cosmetic products, one of which was found to contain three illegal synthetic colorants, namely Basic Blue 26 (0.33 mg/g), Basic Red 2 (0.53 mg/g), and Basic Yellow 28 (31.50 mg/g). Additionally, based on their fragment ions obtained using LC-Q-TOF-MS, the fragmentation pattern of synthetic colorants was predicted. Thus, our work paves the way for the reliable detection of illegal synthetic colorants and may help to prevent the distribution of cosmetics containing the same.
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